A network map of macrophage-stimulating protein (MSP) signaling.
Macrophage-stimulating protein (MSP), a serum-derived growth factor belonging to the plasminogen-related kringle domain family, is mainly produced by the liver and released into the blood. MSP is the only known ligand for RON ("Recepteur d'Origine Nantais", also known as MST1R), which is a member of the receptor tyrosine kinase (RTK) family. MSP is associated with many pathological conditions, including cancer, inflammation, and fibrosis. Activation of the MSP/RON system regulates main downstream signaling pathways, including phosphatidylinositol 3-kinase/ AKT serine/threonine kinase/ (PI3-K/AKT), mitogen-activated protein kinases (MAPK), c-Jun N-terminal kinase (JNK) & Focal adhesion kinase (FAK). These pathways are mainly involved in cell proliferation, survival, migration, invasion, angiogenesis & chemoresistance. In this work, we created a pathway resource of signaling events mediated by MSP/RON considering its contribution to diseases. We provide an integrated pathway reaction map of MSP/RON that is composed of 113 proteins and 26 reactions based on the curation of data from the published literature. The consolidated pathway map of MSP/RON mediated signaling events contains seven molecular associations, 44 enzyme catalysis, 24 activation/inhibition, six translocation events, 38 gene regulation events, and forty-two protein expression events. The MSP/RON signaling pathway map can be freely accessible through the WikiPathways Database URL: https://classic.wikipathways.org/index.php/Pathway:WP5353 .


Prevalence of hearing loss among primary school children in Mbarara, Uganda.
Hearing loss in children is a common entity worldwide. We examined the prevalence and etiology of hearing loss among primary school children in Mbarara, Uganda. Cross-sectional study in primary school children aged 5-14 was performed to determine the prevalence of hearing loss. Ugandan primary school children were screened for disabling hearing loss (threshold >30dB) and confirmatory audiometry was performed on those who failed the screening. There were 639 children screened. Thirty-five (5.5%) of children screened failed and were referred for further testing. Two children were lost to follow-up. The percentage of children with true hearing loss was 3.1%. The incidence of failed hearing screening and hearing loss in Mbarara, Uganda is similar to other populations. Hearing loss is a significant problem in Uganda and efforts should be made for primary, secondary, and tertiary prevention of hearing loss.


Assessment of Mucosa-Associated Epithelial Chemokine, Thymus-Expressed Chemokine, Periostin and Zonulin Levels in Infants With Atopic Dermatitis.
Atopic dermatitis (AD) is a chronic inflammatory skin disease. Skin and gut are the organs that first encounter antigens and environmental triggers. The mechanisms behind the relation between skin and gut immune responses in AD have not been identified yet. To investigate mucosa-associated epithelial chemokine (MEC/CCL28), thymus-expressed chemokine (TECK/CCL25), periostin and zonulin levels in infants with AD. Children under one year old participated in the study. We used a propensity matching score. We included 39 infants who had active AD lesions at the time of evaluation. Serum MEC/CCL28, TECK/CCL25, periostin and zonulin levels were measured. We examined age and sex matched 39 infants with AD and 39 healthy infants. Median value of zonulin was lower in infants with AD [49.2 (27.1-71.8) ng/mL] compared to healthy controls [58.5 (27.3-80.8) ng/mL] (P < 0.001). Infants with zonulin levels ≤55.15 ng/mL had 11.64 times more risk of developing AD than the infants with zonulin levels >55.15 ng/mL. Infants whose MEC/CCL28 levels were ≥8.3 ng/mL had 5.83 times more risk of developing AD than the infants with MEC levels <8.3 ng/mL. Duration of AD and SCORAD index score did not show correlation with MEC/CCL28, TECK/CCL25, periostin and zonulin levels. Low zonulin levels and high MEC/CCL28 levels in infants may show an increased association with AD.


Catastrophizing about their children's pain is related to higher parent-child congruency in pain ratings: an experimental investigation.
Little is known about the variables that account for why parents underestimate the pain of their child. In the present experiment, the joint impact of parental catastrophizing about their child's pain and children's facial pain expressions was examined upon pain estimates of their child undergoing a pressure pain test. In line with previous research, parents underestimated their children's pain. Interestingly, it was found that pain was estimated as higher when the child showed more facial pain expressions and when parents catastrophized more about their child's pain. An intriguing finding was that catastrophizing about their child's pain was related to less parent-child incongruence in pain ratings. The discussion addresses the possible functions of catastrophizing of parents about their children's pain, and delineates avenues for future research.


Renal capsular leiomyoma.
An exophytic renal mass was diagnosed preoperatively as renal cell carcinoma by ultrasound, computerized tomography and arteriography. Intraoperative recognition of the proper diagnosis of leiomyoma of the renal capsule permitted renal-sparing excision. Renal capsular leiomyoma should always be considered in the differential diagnosis of an apparent exophytic renal cell carcinoma.


Experiences of mental health problems vulnerability, psychological symptoms and coping mechanisms of displaced adolescents in North-east Nigeria.
At a global level the issue of forcible internal displacement appears to be somewhat growing. Most internally displaced persons in Nigeria are children and adolescents, but most of the existing literature and public debate focus on the experiences and outcomes of displaced adults. We set out to explore the adverse conditions that increase vulnerability to mental health problems, and the psychological symptoms and coping mechanisms reported by internally displaced adolescents. Using a qualitative approach, 15 adolescents recruited across three different displacement settlements were interviewed individually using a semi-structured interview format on their displacement experiences, and their adaptations to these experiences. The thematic analysis showed that, in addition to experience of mental health problems, vulnerabilities and profound psychological symptoms for some, displacement constituted a complex life-changing process for all. High-risk coping strategies such as 'seeking support by begging' and 'transactional sex for exchange of need were predominantly adopted by female participants. Some participants demonstrated resilience by engaging in religious behavioural coping, which they considered necessary to mitigate their past traumatic memories. Internally displaced adolescents' experience of psychological symptoms and choice of coping mechanisms relate to their contextually perceived needs. Psychosocial support for these adolescents is recommended.


The neuroactive substances and associated muscle system in Rhipidocotyle campanula (Digenea, Bucephalidae) from the intestine of the pike Esox lucius.
We report about the muscular system and the serotonergic and FMRFamidergic components of the nervous system of the Bucephalidae trematode, Rhipidocotyle campanula, an intestinal parasite of the pike. We use immunocytochemical methods and confocal scanning laser microscopy (CLSM). The musculature is identified by histochemical staining with fluorescently labeled phalloidin. The body wall musculature of R. campanula contains three layers of muscle fibres - the outer thin circular, intermediate longitudinal and inner diagonal muscle fibres running in two opposite directions. The digestive system of R. campanula possess of a well-developed musculature: radial, longitudinal and circular muscle elements are detected in the pharynx, circular and longitudinal muscle filaments seen in the oesophagus, and longitudinal and the circular muscle fibres were found in the intestinal wall. Specific staining indicating the presence of actin muscle filaments occurs in the cirrus sac localized in the posterior body region. The frontal region of anterior attachment organ, the rhynchus, in R. campanula is represented by radial muscle fibres. The posterior part of the rhynchus comprise of radial muscles forming the organ's wall, and several strong longitudinal muscle bundles. Serotonergic and FMRFamidergic structures are detected in the central and peripheral compartments of the nervous system of R. campanula, that is, in the paired brain ganglia, the brain commissure, the longitudinal nerve cords, and connective nerve commissures. The innervations of the rhynchus, pharynx, oesophagus and distal regions of the reproductive system by the serotonergic and FMRFamidergic nervous elements are revealed. We compare our findings obtained on R. campanula with related data for other trematodes.


Sterols of the cultured dinoflagellate Pyrocystis lunula.
Eighteen components of the sterol fraction of Pyrocystis lunula have been identified. In addition to 4 alpha-methyl sterols (typical dinoflagellate sterols), regular sterols, both with a saturated and delta 5-unsaturated skeleton, were isolated, together with delta 4-3-keto steroids including the hitherto unknown 23,24R-dimethyl-4,22E-cholestadien-3-one.


T cells recognizing multiple peptides of myelin basic protein are found in blood and enriched in cerebrospinal fluid in optic neuritis and multiple sclerosis.
The cause of multiple sclerosis (MS) is unknown. Recently reported abnormal T-cell responses to several myelin proteins and myelin basic protein (MBP) peptides in peripheral blood constitute one line of evidence that autoimmune mechanisms could be involved in the pathogenesis of the disease. Monosymptomatic unilateral optic neuritis (ON) is a common first manifestation of MS and important to examine for a possible restriction of the T-cell repertoire early in the disease. T-cell activities to MBP and the MBP amino acid sequences 63-88, 110-128 and 148-165 were examined by short-term cultures of mononuclear cells from cerebrospinal fluid (CSF) and blood in the presence of these antigens, and subsequent detection and counting of antigen-specific T cells that responded by interferon-gamma (IFN-gamma) secretion. Most patients with MS and ON had MBP and MBP peptide-reactive T cells in CSF, amounting to mean values of between about 1 per 2000 and 1 per 7000 CSF cells and without immunodominance for any of the peptides. Numbers were 10-fold to 100-fold lower in the patients' blood. Values were similar in ON and MS, and no evidence was obtained for a more restricted T-cell repertoire in ON. The MBP peptide-recognizing T-cell repertoire was different in CSF than in blood in individual patients with ON and MS, thereby giving further evidence for an autonomy of the autoimmune T-cell response in the CSF compartment. No relations were observed between numbers of autoreactive T cells and presence of oligoclonal IgG bands in CSF or abnormalities on magnetic resonance imaging of the brain in ON or clinical variables of MS. The high numbers of MBP and MBP peptide-reactive T cells could play a role in the pathogenesis of ON via secretion of effector molecules, one of them being IFN-gamma, as well as in the transfer of ON to MS.


Novel phleboviruses detected in ticks, Greece.
Since 2009, when severe fever with thrombocytopenia syndrome virus and Heartland virus have been identified and associated with disease in humans, the interest on tick-borne phleboviruses is increasing rapidly. The aim of the present study was to investigate the presence of tick-borne phleboviruses in Greece and compare them with respective ones detected worldwide. Ticks collected from goats and sheep in 60 sites of 13 regional units of Greece were grouped in pools (1-3 ticks per pool) and tested for the presence of phleboviral RNA. Six of 210 pools were positive; they consisted of Rhipicephalus sanguineus ticks collected from sheep in 3 regional units of Greece: Pella (2/30, 6.7%), Imathia (2/21, 9.5%), and Ioannina (2/28, 7.1%). The overall tick minimum infection rate was 2.1%. The sequences of the Greek phlebovirus (provisionally named Antigone virus) form a distinct clade in the tick-borne phleboviruses, differing by >40% from the currently known phleboviruses. Any probable implication of these viruses to public health remains to be elucidated.


Structural basis of TRPC4 regulation by calmodulin and pharmacological agents.
Canonical transient receptor potential channels (TRPC) are involved in receptor-operated and/or store-operated Ca2+ signaling. Inhibition of TRPCs by small molecules was shown to be promising in treating renal diseases. In cells, the channels are regulated by calmodulin (CaM). Molecular details of both CaM and drug binding have remained elusive so far. Here, we report structures of TRPC4 in complex with three pyridazinone-based inhibitors and CaM. The structures reveal that all the inhibitors bind to the same cavity of the voltage-sensing-like domain and allow us to describe how structural changes from the ligand-binding site can be transmitted to the central ion-conducting pore of TRPC4. CaM binds to the rib helix of TRPC4, which results in the ordering of a previously disordered region, fixing the channel in its closed conformation. This represents a novel CaM-induced regulatory mechanism of canonical TRP channels.


Presynaptic actions of curare and atropine on quantal acetylcholine release at a central synapse of Aplysia.
1. In a cholinergic synaptic couple in the buccal ganglion of Aplysia california, where the synaptic areas are situated close to the somata (500 micron), we were able to control transmitter release by stimulating the cell body of the presynaptic neurone with long depolarizing pulses in the presence of tetrodotoxin (TTX). 2. Statistical analysis of noise occurring at the peak of the long-depolarization-induced post-synaptic current (p.s.c.) responses allowed us to calculate the amplitude and the decay time of the miniature post-synaptic currents (m.p.s.c.s). These data were used to calculate the quantal content of the responses. 3. Bath-applied tubocurarine reduced the amplitude of the long-depolarization-induced p.s.c. more than that of the m.p.s.c.s, indicating that tubocurarine exerts a depressive presynaptic action on the quantal content of the post-synaptic responses. 4. Tubocurarine injected into the presynaptic neurone blocked synaptic transmission without decreasing the size of the m.p.s.c.s probably by acting on the mechanism of transmitter release. 5. Bath-applied atropine (10(-6) and 10(-5) M) caused a slight decrease of the m.p.s.c.s but the long-depolarization-induced p.s.c.s increased, as did the quantal content. Higher concentrations of atropine depressed strongly both the m.p.s.c. and the quantal content. 6. Injection of atropine into the presynaptic neurone had the same effect as its bath application, probably due to the leakage of the drug into the synaptic cleft; the effect depended on the concentration reached in the cleft, i.e. on the quantity of injected drug. The synapses of the neighbouring cholinergic neurone were also affected by this leak of atropine. 7. The presence of nicotinic presynaptic receptors blocked by tubocurarine, and muscarinic presynaptic receptors blocked by atropine, which regulate synaptic transmission by facilitating and depressing the ACh release respectively, is discussed.


Catheter application, insertion vein and length of ICU stay prior to insertion affect the risk of catheter-related bloodstream infection.
The Dutch PREZIES surveillance scheme for catheter-related bloodstream infection (CR-BSI) collects data on infection rates and related risk factors. To evaluate risk factors for CR-BSI. Hospitals collected data for intensive care units (ICU) or for the entire hospital. All short-term central venous catheters (CVC), including Swan-Ganz catheters, present for ≥48h were surveyed, except in cases when bacteraemia was present at insertion. CVCs were monitored until infection, removal or death for up to 28 days. Data were collected on 3750 CVCs and 29,003 CVC-days. Of the CVCs surveyed, 1.6% [95% confidence interval (CI) 1.2-2.0] resulted in CR-BSI, representing 2.0/1000 CVC-days (95% CI 1.6-2.6). Multi-variate analysis revealed that the length of ICU stay prior to CVC insertion, insertion in the jugular or femoral vein, and use of the CVC to deliver total parenteral nutrition increased the risk of CR-BSI, whereas use of the CVC to deliver antibiotics decreased the risk of CR-BSI. Attention to these risks has the potential to reduce the incidence of CR-BSI.


Features of a truxene-based stationary phase in capillary gas chromatography for separation of some challenging isomers.
Herein we report the first example of exploring truxene-based derivatives for gas chromatographic (GC) separations. The fabricated thiophene-functionalized truxene (TFT) column exhibited weak polarity and efficiencies as high as 4000plates/m for 0.250mm i.d. columns. TFT column showed preferential retention for halogenated and alkyl benzene analytes, and especially, high resolving capability for the xylene isomers, di- and trichlorobiphenyls (di-CB and tri-CB) isomers. Interestingly, its unique retentions for the latter analytes were found to be closely related with their dihedral angles and the locations of chlorine atoms. This finding on the retention trend has not been reported in GC separations, which may provide a new perspective in elucidating retention behaviours for specific analytes. Moreover, TFT column exhibited high thermal stability up to 320°C and excellent repeatability. This work demonstrates the promising future of truxene derivatives in the separation science.


LINC01468 drives NAFLD-HCC progression through CUL4A-linked degradation of SHIP2.
Accumulating evidence suggests that long noncoding RNAs (lncRNAs) are deregulated in hepatocellular carcinoma (HCC) and play a role in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). However, the current understanding of the role of lncRNAs in NAFLD-associated HCC is limited. In this study, transcriptomic profiling analysis of three paired human liver samples from patients with NAFLD-driven HCC and adjacent samples showed that LINC01468 expression was significantly upregulated. In vitro and in vivo gain- and loss-of-function experiments showed that LINC01468 promotes the proliferation of HCC cells through lipogenesis. Mechanistically, LINC01468 binds SHIP2 and promotes cullin 4 A (CUL4A)-linked ubiquitin degradation, thereby activating the PI3K/AKT/mTOR signaling pathway, resulting in the promotion of de novo lipid biosynthesis and HCC progression. Importantly, the SHIP2 inhibitor reversed the sorafenib resistance induced by LINC01468 overexpression. Moreover, ALKBH5-mediated N6-methyladenosine (m6A) modification led to stabilization and upregulation of LINC01468 RNA. Taken together, the findings indicated a novel mechanism by which LINC01468-mediated lipogenesis promotes HCC progression through CUL4A-linked degradation of SHIP2. LINC01468 acts as a driver of HCC progression from NAFLD, highlights the potential of the LINC01468-SHIP2 axis as a therapeutic target for HCC.


Saddle coronary embolism precipitates flash pulmonary edema.
Coronary artery embolism is as an infrequent but important cause of acute myocardial infarction. We present a rare case of a large saddle embolism in the coronary vasculature which led to an acute myocardial infarction and consequently flash pulmonary edema. Despite intensive medical management, our patient did not survive.


Microbial invasions in sludge anaerobic digesters.
Among processes that control microbial community assembly, microbial invasion has received little attention until recently, especially in the field of anaerobic digestion. However, knowledge of the principles regulating the taxonomic and functional stability of microbial communities is key to truly develop better predictive models and effective management strategies for the anaerobic digestion process. To date, available studies focus on microbial invasions in digesters feed with activated sludge from municipal wastewater treatment plants. Herein, this review summarizes the importance of invasions for anaerobic digestion management, the ecological theories about microbial invasions, the traits of activated sludge microorganisms entering the digesters, and the resident communities of anaerobic reactors that are relevant for invasions and the current knowledge about the success and impacts of invasions, and discusses the research needs on this topic. The initial data indicate that the impact of invasions is low and only a small percentage of the mostly aerobic microorganisms present in the activated sludge feed are able to become stablished in the anaerobic digesters. However, there are still numerous unknowns about microbial invasions in anaerobic digestion including the influence of anaerobic feedstocks or process perturbances that new approaches on microbial ecology could unveil. KEY POINTS: • Microbial invasions are key processes to develop better strategies for digesters management. • Knowledge on pathogen invasions can improve anaerobic digestion microbial safety. • To date, the number of successful invasions on anaerobic digesters from activated sludge organisms is low. • Feed organisms detected in digesters are mostly inactive residual populations. • Need to expand the range of invaders and operational scenarios studied.


The growth and reproduction performance of TALEN-mediated β-lactoglobulin-knockout bucks.
With the technological development of several engineered endonucleases (EENs), such as zinc-finger nucleases, transcription activator-like effector nucleases (TALENs) and CRISPR/Cas9, gene targeting by homologous recombination has been efficiently improved to generate site-specifically genetically modified livestock. However, few studies have been done to investigate the health and fertility of these animals. The purpose of the present study is to investigate if gene targeting events and a recloning procedure would affect the production traits of EEN-mediated gene targeted bucks. TALEN-mediated β-lactoglobulin (BLG) gene mono-allelic knockout (BLG (+/-)) goats and bi-allelic knockout (BLG (-/-)) buck produced by using sequential gene targeting combined with recloning in fibroblasts from BLG (+/-) buck were used to evaluate their health and fertility. Birth weight and postnatal growth of BLG (+/-) bucks were similar to the wild-type goats. None of the parameters for both fresh and frozen-thawed semen quality were significantly different in BLG (+/-) or BLG (-/-) bucks compared to their corresponding comparators. In vitro fertilization (IVF) test revealed that the proportion of IVF oocytes developing to the blastocyst stage was identical among BLG (+/-), BLG (-/-) and wild-type bucks. Conception rates of artificial insemination were respectively 42.3, 38.0 and 42.6 % for frozen-thawed semen from the BLG (+/-), BLG (-/-) and wild-type bucks. In addition, germline transmission of the targeted BLG modification was in accordance with Mendelian rules. These results demonstrated that the analyzed growth and reproductive traits were not impacted by targeting BLG gene and recloning, implicating the potential for dairy goat breeding of BLG (+/-) and BLG (-/-) bucks.


[Xanthogranuloma in adults. Clinico-pathological study of a case].
The authors report a case of naevoxanthoendothelioma in a 58-year-old woman. 24 cases of naevoxanthoendothelioma in adults have already been reported in the literature. If the age of the patient is considered, nosologic problems may arise, especially with the strictly cutaneous reticulohistiocytosis of Senear and Caro. Biological and ultrastructural features allow the authors to develop some hypotheses about pathogenesis of the disease.


PEEK and CFR-PEEK as alternative bearing materials to UHMWPE in a fixed bearing total knee replacement: An experimental wear study.
New bearing materials for total joint replacement have been explored as the need to improve longevity and enhance performance is driven by the changing demands of the patient demographic. Carbon-reinforced PEEK has demonstrated good wear characteristics in experimental wear simulation in both simple geometry pin-on-plate studies and in total hip joint replacement. Carbon reinforced PEEK CFR-PEEK has the potential to reduce tibial insert thickness and preserve bone in the knee. This study investigated the wear performance of PEEK and CFR-PEEK in a low conformity total knee replacement configuration. Custom-made flat inserts were tested against cobalt-chromium femoral bearings in a knee wear simulation for a period of three million cycles. Wear was assessed gravimetrically at intervals throughout the study. The wear rates of both PEEK and CFR-PEEK were very high and almost two orders of magnitude higher than the wear rate of UHMWPE under comparable conditions. Evidence of mechanical failure of the materials, including surface cracking and delamination was observed in both materials. This study highlights that these materials may not be suitable alternatives for UHMWPE in low-conformity designs.


A Detailed History of Retromuscular Repairs for Ventral Hernias: A Story of Surgical Innovation.
We performed a historical review of events concerning retromuscular hernia repairs over the last two centuries. This may shed light on surgical innovators and their novel techniques that have evolved into current practices. Literature reviews of notable surgeons in the subspecialty were reviewed. Historical context was obtained by personal communication with contemporary surgeons who witnessed changes in established techniques firsthand. Even though retromuscular repairs are the central theme of this exercise, it is important to note several adjacent events which steered surgical progress. The status of hernia surgery today is the result of the work of several pioneers separated by time and distance. It may be important to understand the circumstances that have propelled past surgical breakthroughs to stimulate future progress.


Oculodento-osseous dysplasia: heterogeneity or variable expression?
Oculodento-osseous dysplasia (ODOD) has been recognised in three South African patients from two kindreds of Dutch descent. Their ocular, nasal, dental and digital stigmata resembled those of previously reported cases, but their cranial hyperostosis and mandibular overgrowth were of much greater degree. In addition, the two survivors had serious neurological complications consequent upon spinal cord compression at the base of the skull and calcification of the basal ganglia. Two of the patients were the product of marriages between a pair of brothers and a pair of sisters, who were themselves clinically normal. This situation is best explained by autosomal recessive inheritance. As ODOD is usually transmitted as an autosomal dominant, and in view of the unusual severity of the manifestations in our patients, it is possible that the condition is heterogeneous and that they had a distinct autosomal recessive form of the disorder. The presence of minimal stigmata in the mother and two prior generations of our third patient could be equally well interpreted as representing great variation in phenotypic expression of a single dominant gene or manifestation in a heterozygote for a recessive gene.


Biliary obstruction caused by intrabiliary transplantation from hepatocellular carcinoma.
A rare autopsy case of hepatocellular carcinoma (HCC) presenting as extrahepatic bile duct obstruction is reported. A 54-year-old man who had been treated at another hospital for obstructive jaundice was referred to our hospital and admitted on March 1, 1998, because of progressive jaundice. On hospital day 94, he died of bleeding esophageal varices. At autopsy, a bile duct tumor, measuring 3.0 x 3.5 cm and adhering to the wall of the left hepatic duct, occluded the common hepatic duct at the hilus. A tumor measuring 2.0 x 2.0 cm was found in the parenchyma of the left liver lobe. The parenchymal tumor was not continuous with the extrahepatic bile duct tumor. Histologically, the bile duct tumor and the parenchymal tumor of the left lobe were diagnosed as HCC. The bile duct tumor was attached to the mucosa of the bile duct with a thin stalk. No invasive growth into the submucosa was observed. The tumor may have been an intrabiliary transplantation from the HCC in the left lobe via the bile duct.


Vessel density and choroidal vascularity index in patients with Bietti crystalline dystrophy and retinitis pigmentosa.
To evaluate and compare the vessel density (VD) using swept-source optical coherence tomography angiography (OCT-A) and the choroidal vascularity index (CVI) using spectral-domain optical coherence tomography (SD-OCT) in patients with Bietti crystalline dystrophy (BCD) and retinitis pigmentosa (RP). A cross-sectional retrospective study was conducted on 26 eyes of 13 BCD patients, 26 eyes of 13 RP patients, and 26 eyes of 13 age- and gender-matched healthy individuals. BCD patients were further staged as having early, intermediate, and advanced disease. VD was assessed in five quadrants of the macula (superior, temporal, inferior, nasal, and center) using a modified ETDRS technique with OCT-A. SD-OCT scans were binarized using Niblack's autolocal threshold, and CVI was determined as the ratio of the luminal area to the total choroidal area. A significant difference was found in VD in all quadrants of the superficial capillary plexus (SCP) and the deep capillary plexus (DCP) slabs among the three groups (p < 0.001). A statistically significant difference was noted in the mean VD of temporal and inferior quadrants of the SCP and between the BCD and RP groups (p = 0.005, p = 0.015, respectively). A statistically significant difference was observed in the mean VD of the temporal, inferior, and nasal quadrants between the BCD and RP groups on DCP slabs (p = 0.002, p = 0.003, p = 0.003, respectively). The mean central choroidal thickness was 214.65±87.10 μm in the BCD group, 351.69±67.94 μm in the RP group, and 320.92±59.26 μm in the control group (p < 0.001). We found that CVI was significantly higher in the control group than BCD group (p < 0.001), and it was significantly lower in the BCD group when compared to the RP group (p < 0.001).There was no difference in CVI between RP and control groups (p = 0.948). Furthermore, the CVI was significantly lower in the intermediate and advanced disease stages than the early disease stage in the subgroup analysis of BCD patients (p < 0.001, p < 0.001, respectively). CVI is a novel investigative tool to monitor disease progression. The CVI value was lower in BCD and RP patients than in the healthy subjects, and lower CVI values seem to be related to the disease severity in BCD patients. VD was also significantly lower in BCD patients when compared to RP patients, and VD analysis may help clinicians better understand the disease pathophysiology.


[Possibilities for the in vitro evaluation of bioavailability. 5. Compact peroral agents with basic drugs].
Lipophilic, basic drugs can be transported across lipoid-membrane even if the part of the substance being not ionized in the solution is very low. Therefore the resorption model can be used for the control of drugs containing the substances as salts. In the case of 4 substances no differences could be found concerning half transport time between the aqueous solutions and the drugs.


Clinicopathological staging of gastric cancer.
13 175 patients with gastric carcinomas have been staged with reference to the TNM staging system. This has confirmed the poor overall prognosis (5.5 per cent and 3.8 per cent age-adjusted survival rates at 5 and 10 years respectively). However, it has demonstrated the significant survival associated with the treatment of Stage I disease (70.0 per cent at 5 years and 63.2 per cent at 10 years). This study demonstrates that 72.9 per cent of cases are unresected and that interpretation of resectability rates must be related to the population base of the study.


Retrieval of glycoside hydrolase family 9 cellulase genes from environmental DNA by metagenomic gene specific multi-primer PCR.
A new method, termed metagenomic gene specific multi-primer PCR (MGSM-PCR), is presented that uses multiple gene specific primers derived from an isolated gene from a constructed metagenomic library rather than degenerate primers designed based on a known enzyme family. The utility of MGSM-PCR was shown by applying it to search for homologues of the glycoside hydrolase family 9 cellulase in metagenomic DNA. The success of the multiplex PCR was verified by visualizing products on an agarose gel following gel electrophoresis. A total of 127 homologous genes were amplified with combinatorial multi-primer reactions from 34 soil DNA samples. Multiple alignments revealed extensive sequence diversity among these captured sequences with sequence identity varying from 26 to 99.7%. These results indicated that significantly diverse homologous genes were indeed readily accessible when using multiple metagenomic gene specific primers.


Transplantation of the lungs.
Over the last eight years a completely new form of treatment has been introduced for end-stage pulmonary vascular disease and chronic lung disease, including cystic fibrosis. Lung transplantation and heart-lung transplantation have moved from an experimental and innovative stage to clinical treatments. The exclusion criteria for selecting potential recipients have become relaxed whilst preservation techniques have improved to allow ischaemic times of donor organs to be extended beyond 4 h. Monitoring of graft function with daily spirometry and use of transbronchial lung biopsy have enabled early diagnosis of lung rejection or infection and thus effective treatment. Lung transplantation and heart-lung transplantation, by their success, are providing new insight into lung disease and pulmonary physiology.


An update of COVID-19 influence on waste management.
COVID-19 has been sweeping the world. The overall number of infected persons has been increased from 5 M in March 2020 to over 22 M in August 2020 and growing, which seems not to get its peak at the current stage. This has contributed to waste generation and different phases of challenges in waste management practices. The impacts including change in waste amount, composition, timing/frequency (temporal), distribution (spatial) and risk, which affects the handling and treatment practices. Recent impacts, challenges and developments on waste management in the response of COVID-19 have been assessed in this update. Singapore, the cities of Shanghai in China and Brno in the Czech Republic (a member state of the European Union), representing different pandemic development situation and also various cultural attitudes, are specifically analysed and discussed with current data. However, it should be noted that it is still fast developing. A varying trend in term of the waste amount is identified. Shanghai is showing a ~23% decline in household waste amount; however, Singapore is showing a ~3% increase, and Brno is showing a ~1% increase in household waste amount but ~40% decline in business and industrial waste. Manual sorting and recycling have been reported as restricted due to safety precaution. This is supported by the interview communication with ZEVO SAKO (the largest incineration plant in the Czech Republic). This study highlighted that the practices or measures at each place could serve as a guideline and reference. However, adaption is required according to the geographical and socioeconomic factors.


Detailed study of post-Chernobyl Cs-137 redistribution in the soils of a small agricultural catchment (Tula region, Russia).
A detailed study of 137Cs redistribution was conducted within a small agricultural catchment in the highly contaminated Plavsk radioactive hotspot in the Tula region of Central Russia, 32 years after the Chernobyl nuclear power plant (NPP) accident, which occurred on April 26, 1986. Although more than three decades have passed since the Chernobyl NPP incident, 137Cs contamination is high. The 137Cs inventory varies from 67 to 306 kBq·m-2, which is 2-6 times higher than the radiation safety standard; however, the soils remain suitable for crop cultivation. The initial 137Cs fallout within the Plavsk radioactive hotspot was extremely heterogeneous, with a trend of decreasing 137Cs inventories from the NW to the SE directions within the studied territory. Contemporary 137Cs inventories are also very heterogeneous in the studied catchment. However, the trend of the initial 137Cs fallout does not appear in the contemporary 137Cs inventories on the slopes. Two methods of interpolation (expert-visual and automatic) were used to calculate the 137Cs budget, revealing high similarity in their 137Cs loss estimates; however, a large discrepancy was observed in their 137Cs gain estimates. A detailed analysis of 137Cs redistribution revealed the importance of hollows and "plow ramparts" (positive topographic forms on the boundaries of cultivated fields) in the transport and deposition of sediments. A quarter of the total 137Cs gain was deposited within the arable land, whereas a quarter was deposited within the non-plowing sides of the dry valley; the other half was deposited in the valley bottom. About 7-8 × 106 kBq of the 137Cs inventory flowed out of the catchment area, which was only about 2% of the 137Cs fallout after the Chernobyl NPP accident. About 89% of the total 137Cs reserve is concentrated in the top (0-25 cm) layer of soils, regardless of land use or location within the catchment.


Studying kinetochore-fiber ultrastructure using correlative light-electron microscopy.
Electron microscopy (EM) has dominated high-resolution cellular imaging for over 50 years, thanks to its ability to resolve on nanometer-scale intracellular structures such as the microtubules of the mitotic spindle. It is advantageous to view the cell of interest prior to processing the sample for EM. Correlative light-electron microscopy (CLEM) is a technique that allows one to visualize cells of interest by light microscopy (LM) before being transferred to EM for ultrastructural examination. Here, we describe how CLEM can be applied as an effective tool to study the spindle apparatus of mitotic cells. This approach allows transfected cells of interest, in desirable stages of mitosis, to be followed from LM to EM. CLEM has often been considered as a technically challenging and laborious technique. In this chapter, we provide step-by-step pictorial guides that allow successful CLEM to be achieved. In addition, we explain how it is possible to vary the sectioning plane, allowing spindles and microtubules to be analyzed from different angles, and the outputs that can be obtained from these methods when applied to the study of kinetochore fiber ultrastructure.


"It's probably an STI because you're gay": a qualitative study of diagnostic error experiences in sexual and gender minority individuals.
There is a critical need to identify specific causes of and tailored solutions to diagnostic error in sexual and gender minority (SGM) populations. To identify challenges to diagnosis in SGM adults, understand the impacts of patient-reported diagnostic errors on patients' lives and elicit solutions. Qualitative study using in-depth semistructured interviews. Participants were recruited using convenience and snowball sampling. Recruitment efforts targeted 22 SGM-focused organisations, academic centres and clinics across the USA. Participants were encouraged to share study details with personal contacts. Interviews were analysed using codebook thematic analysis. Interviewees (n=20) ranged from 20 to 60 years of age with diverse mental and physical health symptoms. All participants identified as sexual minorities, gender minorities or both. Thematic analysis revealed challenges to diagnosis. Provider-level challenges included pathologisation of SGM identity; dismissal of symptoms due to anti-SGM bias; communication failures due to providers being distracted by SGM identity and enforcement of cis-heteronormative assumptions. Patient-level challenges included internalised shame and stigma. Intersectional challenges included biases around factors like race and age. Patient-reported diagnostic error led to worsening relationships with providers, worsened mental and physical health and increased self-advocacy and community-activism. Solutions to reduce diagnostic disparities included SGM-specific medical education and provider training, using inclusive language, asking questions, avoiding assumptions, encouraging diagnostic coproduction, upholding high care standards and ethics, involving SGM individuals in healthcare improvement and increasing research on SGM health. Anti-SGM bias, queerphobia, lack of provider training and heteronormative attitudes hinder diagnostic decision-making and communication. As a result, SGM patients report significant harms. Solutions to mitigate diagnostic disparities require an intersectional approach that considers patients' gender identity, sexual orientation, race, age, economic status and system-level changes.


Interactive media for parental education on managing children chronic condition: a systematic review of the literature.
Although some research has examined the use of games for the education of pediatric patients, the use of technology for parental education seems like an appropriate application as it has been a part of the popular culture for at least 30 years. The main objective of this systematic review is to examine the literature for research evaluating the use of interactive media in the education of parents of children with chronic conditions. We searched the MEDLINE, PSYCHINFO, CINAHL, Cochrane database of systematic reviews and EMBASE databases from 1986 to 2014 seeking original investigations on the use of interactive media and video games to educate parents of children with chronic conditions. Cohort studies, randomized control trials, and observational studies were included in our search of the literature. Two investigators reviewed abstracts and full texts as necessary. The quality of the studies was assessed using the GRADE guidelines. Overall trend in the results and the degree of certainty in the results were considered when assessing the body of literature pertaining to our focused questions. Our initial search identified 4367 papers, but only 12 fulfilled the criterion established for final analysis, with the majority of the studies having flaws that reduced their quality. These papers reported mostly positive results supporting the idea that parent education is possible through interactive media. We found limited evidence of the effectiveness of using serious games and or interactive media to educate parents of children with chronic conditions.


Prevalence of disturbed eating behavior and associated symptoms of anxiety and depression among adult males and females with type 1 diabetes.
The increased prevalence of disturbed eating behaviors (DEB), depression, and anxiety in type 1 diabetes (T1D) is generally well established; however the majority of existing research to date has focused on female adolescents and young adults. Data on males and older females is scarce. The aim of this study was to assess prevalence of DEB and symptoms of depression and anxiety among adult males and females with type 1 diabetes, to investigate differences between individuals scoring below and above the cut-off on psychopathology, and to examine patterns of eating disorder psychopathology by age and weight. A total of 282 adults with type 1 diabetes aged 18-79 years participated in the study. Measures included the Diabetes Eating Problem Survey - Revised (DEPS-R), the Hospital Anxiety and Depression Scale (HADS), and clinical data from the Norwegian Quality Improvement of Laboratory Examinations (NOKLUS) system. A total of 20.3% of the whole sample (13.3% among males and 24.8% among females) scored above the DEPS-R cut-off score for DEB. As for depression and anxiety, the prevalence in the whole sample was 6.2% and 19.0%, respectively. The prevalence was generally higher in females than males across all psychopathology measures. HbA1c was significantly associated with the DEPS-R total score (p < .01) among females, but not with depression and anxiety. Mean DEPS-R score decreased with increasing age, and when our previous reported data from children and adolescents are included, a peak prevalence in DEB in adolescence and young adult age is demonstrated. The results of this study point to the need for increased awareness of psychological comorbidity among adults with type 1 diabetes, in particular young adult females. Screening is recommended to secure early detection and subsequent intervention for these individuals.


Developmental toxicity of halogenated acetonitriles: drinking water by-products of chlorine disinfection.
The developmental toxicity of acetonitrile and 5 halogenated derivatives was examined with an in vivo teratology screen adapted for use in the Long-Evans rat. The screen was extended to an evaluation of growth till postnatal Days 41-42, and weight of several organs at sacrifice. Acetonitrile was without developmental effects even at doses toxic to the dam. Of the halogenated compounds, treatment with trichloroacetonitrile (TCAN) and dichloroacetonitrile (DCAN) resulted in reduced fertility and increased early implantation failure. There was no effect on litter size in females bearing live litters, but pup birth weight was reduced in all litters exposed to halogenated compounds. Perinatal survival of the pups was adversely impacted by DCAN and TCAN. Postnatal growth till Day 4 was reduced by DCAN and bromochloroacetonitrile (BCAN) while growth till Day 42 was consistently affected only by TCAN. Some general observations were made on the usefulness of the criteria used in the screen, and TCAN, the most toxic of the halogenated compounds, was selected for further in-depth evaluation.


Synergistic stimulation of osteoblast differentiation of rat mesenchymal stem cells by leptin and 25(OH)D3 is mediated by inhibition of chaperone-mediated autophagy.
Vitamin D is important for the mineralization of bones by stimulating osteoblast differentiation of bone marrow mesenchymal stem cells (BMMSCs). BMMSCs are a target of vitamin D action, and the metabolism of 25(OH)D3 to biologically active 1α,25(OH)2D3 in BMMSCs promotes osteoblastogenesis in an autocrine/paracrine manner. Our previous study with human BMMSCs showed that megalin is required for the 25(OH)D3-DBP complex to enter cells and for 25(OH)D3 to stimulate osteoblast differentiation in BMMSCs. Furthermore, we reported that leptin up-regulates megalin in those cells. Leptin is a known inhibitor of PI3K/AKT-dependent chaperone-mediated autophagy (CMA). In this study, we tested the hypothesis that leptin acts synergistically with 25(OH)D3 to promote osteoblastogenesis in rat BMMSCs by a mechanism that entails inhibition of PI3K/AKT-dependent CMA. BMMSCs were isolated from rat bone marrow (4-week-old male SD rats); qRT-PCR and western immunoblots or immunofluorescence were used to evaluate the expression of megalin, ALP, COL1A1, RUNX2, OSX, OSP, and CMA in rBMMSCs. The osteoblast differentiation was evaluated by ALP activity, ALP staining, and calcium deposition. The viability of rBMMSCs was assessed with the CCK-8 kit. Biosynthesis of 1α,25(OH)2D3 was measured by a Rat 1α,25(OH)2D3 ELISA Kit. The combination of leptin and 25(OH)D3 treatment significantly enhanced osteoblast differentiation as shown by ALP activity, ALP staining, and calcium deposition, the expression of osteogenic genes ALP, COL1A1, RUNX2, OSX, and OSP by qRT-PCR and western immunoblots in rBMMSCs. Leptin enhanced the expression of megalin and synthesis of 1α,25(OH)2D3 in rBMMSCs. Our data showed that leptin inhibited CMA activity of rBMMSCs by activating PI3K/AKT signal pathway; the ability of leptin to enhance 25(OH)D3 promoted osteoblast differentiation of rBMMSCs was weakened by the PI3K/AKT signal pathway inhibitor. Our data reveal the mechanism by which leptin and 25(OH)D3 promote osteoblast differentiation in rBMMSCs. Leptin promoted the expression of megalin by inhibiting CMA, increased the utilization of 25(OH)D3 by rBMMSCs, and enhanced the ability of 25(OH)D3 to induce osteoblast differentiation of rBMMSCs. PI3K/AKT is at least partially involved in the regulation of CMA. These data indicate the importance of megalin in BMMSCs for vitamin D's role in skeletal health.


PTEN deficiency facilitates the therapeutic vulnerability to proteasome inhibitor bortezomib in gallbladder cancer.
Gallbladder cancer (GBC) is an aggressive malignancy of biliary tract with poor prognosis. Although several studies have shown the frequency of relevant genetic alterations, there are few genetic models or translational studies that really benefit for GBC treatment in the era of precision medicine. By targeted sequencing and immunohistochemistry staining, we identified that phosphate and tension homology deleted on chromosome ten (PTEN) was frequently altered in GBC specimens, and loss of PTEN expression was independently correlated with poor survival outcomes. Further drug screening assays revealed proteasome inhibitor bortezomib as a promising agent for GBC treatment, and knockdown of PTEN increased bortezomib efficacy both in vivo and in vitro. Therapeutic evaluation of patient derived xenografts (PDXs) strongly supported the utilization of bortezomib in PTEN deficient GBC. Mechanically, functional PTEN inhibited ARE-dependent transcriptional activity, the same machinery regulating the transcription of proteasome subunits, thus PTEN suppressed proteasome activity and bortezomib sensitivity. Through siRNA screening, we identified the ARE-related transcriptional suppressor BACH1 involved in PTEN-mediated proteasome inhibition and regulated by PTEN-AKT1 axis. In summary, our study indicates that proteasome activity represents a prime therapeutic target in PTEN-deficient GBC tumors, which is worthy of further clinical validation.


2-Aminopurine hairpin probes for the detection of ultraviolet-induced DNA damage.
Nucleic acid exposure to radiation and chemical insults leads to damage and disease. Thus, detection and understanding DNA damage is important for elucidating molecular mechanisms of disease. However, current methods of DNA damage detection are either time-consuming, destroy the sample, or are too specific to be used for generic detection of damage. In this paper, we develop a fluorescence sensor of 2-aminopurine (2AP), a fluorescent analogue of adenine, incorporated in the loop of a hairpin probe for the quantification of ultraviolet (UV) C-induced nucleic acid damage. Our results show that the selectivity of the 2AP hairpin probe to UV-induced nucleic acid damage is comparable to molecular beacon (MB) probes of DNA damage. The calibration curve for the 2AP hairpin probe shows good linearity (R(2)=0.98) with a limit of detection of 17.2 nM. This probe is a simple, fast and economic fluorescence sensor for the quantification of UV-induced damage in DNA.


An ortho-Quinone Methide Mediates Disulfide Migration in the Biosynthesis of Epidithiodiketopiperazines.
The transannular disulfide functions as a key structural element imparting diverse biological activities to epidithiodiketopiperazines (ETPs). Although mechanisms were proposed in previous studies, α,β'-disulfide formation in ETPs is not well-determined owing to the failure to identify the hypothetical intermediate. Herein, we characterize the key ortho-quinone methide (o-QM) intermediate and prove its involvement in the carbon-sulfur migration from an α,α'- to an α,β'-disulfide by elucidating pretrichodermamide A biosynthesis, which is catalyzed by a FAD-dependent thioredoxin oxygenase TdaE harboring a noncanonical CXXQ motif. Biochemical investigations of recombinant TdaE and mutants demonstrated that the construction of the α,β'-disulfide was initiated by Gln140 triggering proton abstraction for generation of the essential o-QM intermediate, accompanied by β'-acetoxy elimination. Subsequent attack on the α,α'-disulfide by Cys137 led to disulfide migration and spirofuran formation. This study expands the biocatalytic toolbox for transannular disulfide formation and sets the stage for the targeted discovery of bioactive ETPs.


EEG differences as a function of creativity, stage of the creative process, and effort to be original.
Results of two experiments examining the relationship between creativity and EEG alpha wave presence are reported. In Experiment 1, it was found that more creative subjects exhibited higher alpha indices during an analogue of creative inspiration than during an analogue of creative elaboration. This pattern was not found in less creative subjects. In Experiment 2 a similar effect was found in a more controlled setting and shown to be specific to creative subjects when they are instructed to be original but not when they are given no such instructions. No consistent relationship between creativity and basal EEG alpha activity was found.


ADNet++: A few-shot learning framework for multi-class medical image volume segmentation with uncertainty-guided feature refinement.
A major barrier to applying deep segmentation models in the medical domain is their typical data-hungry nature, requiring experts to collect and label large amounts of data for training. As a reaction, prototypical few-shot segmentation (FSS) models have recently gained traction as data-efficient alternatives. Nevertheless, despite the recent progress of these models, they still have some essential shortcomings that must be addressed. In this work, we focus on three of these shortcomings: (i) the lack of uncertainty estimation, (ii) the lack of a guiding mechanism to help locate edges and encourage spatial consistency in the segmentation maps, and (iii) the models' inability to do one-step multi-class segmentation. Without modifying or requiring a specific backbone architecture, we propose a modified prototype extraction module that facilitates the computation of uncertainty maps in prototypical FSS models, and show that the resulting maps are useful indicators of the model uncertainty. To improve the segmentation around boundaries and to encourage spatial consistency, we propose a novel feature refinement module that leverages structural information in the input space to help guide the segmentation in the feature space. Furthermore, we demonstrate how uncertainty maps can be used to automatically guide this feature refinement. Finally, to avoid ambiguous voxel predictions that occur when images are segmented class-by-class, we propose a procedure to perform one-step multi-class FSS. The efficiency of our proposed methodology is evaluated on two representative datasets for abdominal organ segmentation (CHAOS dataset and BTCV dataset) and one dataset for cardiac segmentation (MS-CMRSeg dataset). The results show that our proposed methodology significantly (one-sided Wilcoxon signed rank test, p<0.05) improves the baseline, increasing the overall dice score with +5.2, +5.1, and +2.8 percentage points for the CHAOS dataset, the BTCV dataset, and the MS-CMRSeg dataset, respectively.


Characterization of glycerol nonutilizing and protoperithecial mutants of Neurospora.
Mutants defective in polyol metabolism and/or in protoperithecial development were selected in Neurospora tetrasperma, a species in which protoperithecial development occurs at nonpermissively high temperature if certain polyols are used in lieu of sucrose as carbon source. Mutants selected for nonutilization of one of the four polyols tested, glycerol, mannitol, sorbitol, or xylitol, were usually found to be nonutilizers of the other three polyols as well. Mutants blocked at various stages of protoperithecial development complemented pairwise to produce more advanced developmental stages, usually mature protoperithecia and, when of opposite mating type, mature perithecia. About one-third of the mutants manifested both polyol auxotrophy and defective protoperithecial development upon initial isolation, but protoperithecial defectiveness in such mutants usually showed erratic segregation in crosses and/or instability to repeated vegetative transfer, whereas polyol auxotrophy usually did not and was, therefore, studied further. Two glycerol nonutilizing strains were introgressed into N. crassa to facilitate genetic analysis. One, glp-4, lacked both inducible and constitutive glycerol kinase and mapped to linkage group VI, between ad-1 and rib-1; the other, glp-5, lacked glyceraldehyde kinase and mapped to linkage group I, proximal to ad-9. Another mutant, gly-u(234), has been reported by other investigators to lack inducible glycerol kinase but to map to linkage group I, distal to ad-9.


Erratum to: Three-dimensional thoracoscopic vertebral body replacement at the thoracolumbar junction.
Erratum to:Oper Orthop Traumatol 2018 https://doi.org/10.1007/s00064-018-0559-3 The article was wrongly published under the article type "Review". Please note that the article is an "Original Paper".The publisher apologizes to authors and ….


A systematic review of geographic information systems based methods and criteria used for electric vehicle charging station site selection.
Many studies have incorporated particular models with various methods and algorithms to resolve the site selection problem for electric vehicle charging stations (EVCS). This paper systematically reviews research that evaluates geographic information systems (GIS) based EVCS location techniques and the variables used for decision making. We classify and characterize those techniques and variables to determine important linkages within the literature. A variety of databases were referenced to extract research published from 2010 to March 2023 pertinent to this specific location optimization problem, and 74 papers were selected after thorough evaluation. The models used in each paper were examined along with the methods for selecting variables and ranking alternate locations. Site selection for EVCS requires a multi-criteria decision making approach to meet the sustainability, efficiency, and performance goals of communities adopting electric vehicle mobility. Our results indicate that map algebra and data overlay methods have been used more frequently with GIS-based analysis than other techniques, while geographic and demographic variables are commonly the most significant site selection characteristics. The reviewed methods have most often been applied to urban locations; however, the transfer of these techniques to a rural EVCS site selection problem has been rarely explored in the current literature. This research assessment contributes relevant guidance for the application of methodologies useful in policymaking and provides recommendations for future research based on these findings.


Involvement of tRNA in retrovirus expression: biological implications of reverse transcriptase-primer tRNA interactions.
We have previously studied the topographical and functional implications of the recognition of primer tRNATrp by avian retrovirus reverse transcriptase. Here we have presented evidence that the enzyme is able to deacylate beef liver Trp-tRNATrp, provided that 35-S viral RNA is present in the incubation mixture. No effect of dNTPs on this activity was observed. The extensive modification of tRNATrp with acrylonitrile led to a marked loss of priming activity by tRNATrp if the annealing between primer and template was performed at 37 degrees C, while the annealing of cyanoethylated tRNA with the viral genome at 75 degrees C gave almost normal levels of cDNA synthesis. We have also studied the priming behaviour of tRNATrp, modified by incorporation of various analogs of adenosine. Only tRNATrp-2'dA was active in cDNA initiation; 3'dA, 3'NH2-3'dA, and primer tRNA with formycin in the 3' end showed low or nonexistent priming activity.


[Oligo(2'-O-Methylribonucleotides) and their derivatives: IV. Conjugates of oligo(2'-O-methylribonucleotides) with minor groove binders and intercalators: synthesis, properties and application].
Conjugates of pyrimidine triplex forming 3'-protected oligo(2'-O-methylribonucleotides) with minor groove binders (MGB) and triplex specific intercalator benzoindoloquinoline (BIQ) at 5'-terminus were synthesized. The conjugates formed stable complexes with target dsDNA by simultaneous binding both in its minor and major grooves and BIQ intercalation. The dissociation constants and thermal stability of the conjugate complexes with model dsDNA corresponding to polypurine tract (PPT) of genes nef and pol from HIV proviral genome were determined. Conjugation of oligo(2'-O-methylribonucleotides) with MGB and intercalator increased the stability of the triple complexes with dsDNA at pH 7.2 and 37 degrees C. Intercalator introduction accelerates the process of complex formation. Dose-dependent arrest of the in vitro transcription was demonstrated when a 780 b.p. DNA fragment containing the polypurine tract was transcribed under the control of T7 promoter in the presence of different concentrations of conjugates of oligo(2'-O-methylribonucleotides) containing MGB and BIQ intercalator.


Studies on amendment of different biopolymers in sandy loam and their effect on germination, seedling growth of Gossypium herbaceum L.
Different biopolymers, agar, cellulose, alginate, psyllium gaur gum, and bacterial exopolysaccharide (EPS) powders were amended to check their efficacy in enhancing maximum water holding capacity (MWHC), permanent wilting point (PWP), and germination and seedling growth of the Gossypium herbaceum in a laboratory scale. The efficacy of all biopolymers for enhancement of MWHC, PWP, and growth was also analyzed by measuring organic carbon, organic matter, total nitrogen, respiration rate, and microflora in amended and control sandy loams. The range of concentrations (0.2-2%) of all biopolymers was incorporated in sandy loam containing pots. The soil without polymer was considered as control. The psyllium (0.6%) and bacterial EPS (1%) amended soil has 242 and 233% increase in MWHC and thus delaying in the permanent wilting point by 108 and 84 h at 37 °C, respectively, as compared to control. All biopolymers found to increase more or less MWHC, organic matter, total nitrogen, microflora, and PWP as compared to control. The psyllium and bacterial EPS show the highest increase organic matter, biomass, and microflora. The highest reduction in MWHC after 12 weeks were observed in cellulose, gaur gum, and alginate; besides, psyllium, bacterial EPS, and agar showed comparatively less reduction MWHC, i.e., 24% and 14.5%, respectively. The toxicity studies of biopolymer were carried out on earthworm (Eisenia foetida). It revealed their nontoxic nature. The biopolymer amendment in sandy loam can be an effective strategy to improved soil texture, fertility, and thereby crop yield.


Association of Malassezia pachydermatis with systemic infections of humans.
Thirty-two Malassezia spp. isolates from human clinical specimens represented M. furfur and M. pachydermatis. Both species reportedly were obtained from patients with similar febrile systemic syndromes, including infections of the lungs or other tissues.


Acute alcohol consumption-induced let-7a inhibition exacerbates hepatic apoptosis by regulating Rb1 in mice.
Alcohol consumption is a critical risk factor for hepatic pathogenesis, including alcoholic liver diseases (ALD), but implications of alcohol-induced dysregulation of microRNA (miRNA) in ALD pathogenesis are not completely understood. In the present study, C57BL/6J male mice were treated with saline (CON; oral gavage; n = 8) or alcohol (EtOH; 3 g/kg body weight; oral gavage; n = 8) for 7 days. A total of 599 miRNAs and 158 key mRNAs related to fatty liver and hepatotoxicity pathways were assessed in mice liver tissues. The mRNA expression datasets were then utilized to predict interactions with miRNAs that were changed by alcohol consumption. Predicted miRNA-mRNA interactions were validated using in vitro miRNA transfection experiments. The results showed that let-7a was significantly decreased in the EtOH group and Rb1 mRNA was predicted as a target gene. This was further supported by an inverse correlation of RB1 and let-7a expression in mice liver tissue. Additionally, key protein expressions involved in RB1-apoptosis axis [i.e., p73, cleaved CASP-3 (cCASP-3), and cCASP-7] showed a trend of increase in the EtOH mice; this was also confirmed by capase-3 enzyme activity and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay in livers of mice that had consumed alcohol. In line with our in vivo observations, alcohol treatment suppressed the let-7a expression and subsequently upregulated p73, cCASP-3, and cCASP-7 protein expressions in mice hepatocytes. Additional proteins in the apoptosis regulatory pathway (i.e., MDM2-p53 axis) were significantly changed in response to let-7a suppression in the cells. Taken together, the current study provides mechanistic evidence that alcohol consumption-induced let-7a suppression results in the upregulation of RB1, thereby promoting hepatic apoptosis through induction of pro-apoptotic proteins (e.g., p73), and by, at least in part, preventing MDM2-mediated p53 degradation.


Peak urine flow as a predictor of urine infection and retention after coronary artery bypass grafting.
Peak urine flow rates were measured in 83 men before coronary artery bypass grafting. The patients were grouped according to the method of Drach et al. [1] into those with normal, equivocal or abnormal flow rates. No cases of urine infection occurred in any of the 3 groups and only 2 patients in the group with abnormal peak flow developed urine retention. This was not statistically significant. Peak urine flow did not appear to be useful in predicting urine retention after coronary artery bypass grafting.


Identifying long non-coding RNAs and characterizing their functional roles in swine mammary gland from colostrogenesis to lactogenesis.
This study was conducted to identify the functional long non-coding RNAs (lncRNAs) for swine lactation by RNA-seq data of mammary gland. According to the RNA-seq data of swine mammary gland, we screened lncRNAs, performed differential expression analysis, and confirmed the functional lncRNAs for swine lactation by validation of genome wide association study (GWAS) signals, functional annotation and weighted gene co-expression network analysis (WGCNA). We totally identified 286 differentially expressed (DE) lncRNAs in mammary gland at different stages from 14 days prior to (-) parturition to day 1 after (+) parturition, and the expressions of most of lncRNAs were strongly changed from day -2 to day +1. Further, the GWAS signals of sow milk ability trait were significantly enriched in DE lncRNAs. Functional annotation revealed that these DE lncRNAs were mainly involved in mammary gland and lactation developing, milk composition metabolism and colostrum function. By performing weighted WGCNA, we identified 7 out of 12 lncRNA-mRNA modules that were highly associated with the mammary gland at day -14, day -2, and day +1, in which, 35 lncRNAs and 319 mRNAs were involved. This study suggested that 18 lncRNAs and their 20 target genes were promising candidates for swine parturition and colostrum occurrence processes. Our research provided new insights into lncRNA profiles and their regulating mechanisms from colostrogenesis to lactogenesis in swine.


Phytochrome B regulates reactive oxygen signaling during abiotic and biotic stress in plants.
Reactive oxygen species (ROS) and the photoreceptor protein phytochrome B (phyB) play a key role in plant acclimation to stress. However, how phyB that primarily functions in the nuclei impacts ROS signaling mediated by respiratory burst oxidase homolog (RBOH) proteins that reside on the plasma membrane, during stress, is unknown. Arabidopsis thaliana and Oryza sativa mutants, RNA-Seq, bioinformatics, biochemistry, molecular biology, and whole-plant ROS imaging were used to address this question. Here, we reveal that phyB and RBOHs function as part of a key regulatory module that controls apoplastic ROS production, stress-response transcript expression, and plant acclimation in response to excess light stress. We further show that phyB can regulate ROS production during stress even if it is restricted to the cytosol and that phyB, respiratory burst oxidase protein D (RBOHD), and respiratory burst oxidase protein F (RBOHF) coregulate thousands of transcripts in response to light stress. Surprisingly, we found that phyB is also required for ROS accumulation in response to heat, wounding, cold, and bacterial infection. Our findings reveal that phyB plays a canonical role in plant responses to biotic and abiotic stresses, regulating apoplastic ROS production, possibly while at the cytosol, and that phyB and RBOHD/RBOHF function in the same regulatory pathway.


Effect of St. John's wort extract Ze 117 on the lateral mobility of β1-adrenergic receptors in C6 cells.
Depression has been associated with altered signal transduction of serotonergic, dopaminergic and adrenergic neurotransmitter systems in the brain. Signaling relies on receptor-ligand interactions and subsequent regulatory processes, but also on lateral receptor mobility. The aim of this study was to investigate the effect of the St. John's wort extract Ze 117 on the lateral mobility of SNAP-tagged β1-adrenergic receptors (β1AR) in the plasma membrane of C6 cells under both, non-stimulating and isoprenaline-stimulating conditions. Single particle tracking (SPT) was used, whereby the registered trajectories were evaluated by variational Bayesian treatment of a hidden Markov model (vbSPT) and packing coefficient (Pc) analysis with respect to diffusion coefficients, receptor state occupancies and confinement. Three different diffusion states were identified, differing in their diffusion coefficients. Treatment with Ze 117 [25 µg/ml] decreased the mobility of the β1AR, which was manifested by a relative increase in the slow-diffusing state S1 (0.21-0.30) compared to control and by an increase in receptor confinement (79.4-68.1 nm). After isoprenaline stimulation of control cells, the slow-diffusing state was more pronounced, whereas confinement was not affected. In summary, SPT has been shown to be a powerful method to analyze lateral receptor mobility. Furthermore, the present study identified a correlation between Ze 117 treatment and β1AR mobility.


Cystoisospora canis (Apicomplexa: Sarcocystidae): development of monozoic tissue cysts in human cells, demonstration of egress of zoites from tissue cysts, and demonstration of repeat monozoic tissue cyst formation by zoites.
Sporozoites of Cystoisospora canis penetrated and developed to monozoic tissue cysts in 4 human, 1 monkey, 1 bovine and 2 canine cell lines. No asexual division was documented although multiple infection of a single cell was observed. Examination of cultures using transmission electron microscopy demonstrated that they were monozoic tissue cysts and contained a single sporozoite. The appearance of monozoic tissue cysts in all cell lines was similar but the parasitophorous vacuole surrounding some sporozoites in DH82 dog macrophages was swollen. Monozoic tissue cysts were observed for up to 127 days in human pigmented retinal epithelial cells. Treatment of cell cultures containing monozoic tissue cysts with 0.75 sodium taurocholic acid and 0.25% trypsin stimulated egress of zoites (former sporozoites) from tissue cysts. Zoites collected from monozoic tissue cysts were able to penetrate and develop to monozoic tissue cysts in new host cells. Monozoic tissue cysts survived exposure to acid pepsin solution indicating that they would be orally infectious. The tissue cyst wall surrounding zoites did not autofluoresce as did oocyst and sporocyst walls exposed to UV light. We believe that C. canis can be used as a model system to study extra-intestinal monozoic tissue cysts stages of Cystoisospora belli of humans.


Membrane rectification in single smooth muscle cells from the rat tail artery.
Membrane rectification to depolarization was studied by voltage recording with patch electrodes in freshly isolated cells from the rat tail artery. Injection of depolarizing currents elicited electrotonic potentials that developed with a single-exponential time course (time constant of 94.8 ms). When the cell was depolarized beyond -30 mV, delayed rectification was observed. A second type of rectification, characterized by oscillations, was observed when the cell was depolarized positive to +30 mV. The threshold of this rectification and the oscillations were sensitive to changes in intracellular Ca2+. Delayed rectification was more sensitive to 4-aminopyridine but more resistant to tetraethylammonium and charybdotoxin than the Ca(2+)-sensitive rectification. A 4-aminopyridine-sensitive outward current (IK,dr) with a threshold of around -30 mV and a second Ca(2+)-sensitive outward current (IK,Ca) activated at around +30 mV were observed from whole-cell voltage clamp recordings. IK,Ca was blocked by tetraethylammonium and charybdotoxin. An 11-pS and a 122-pS channel, having characteristics similar to IK,dr and IK,Ca respectively, were identified from single-channel recordings. These observations showed how membrane depolarization of vascular smooth-muscle cells was regulated by these two populations of K+ channels under various conditions.


Further findings on assaultiveness and alcohol use in interpersonal disputes.
In a previous study uniform observational data were collected regarding family disputes managed by police officers; those data contradicted views commonly held by social scientists and by police about the roles of assaultiveness and alcohol use in these events. In the present study police officers employed systematic naturalistic observation during their interventions as third parties in disputes between nonfamily members as well as those between family members. Tentative conclusions about assaultiveness, based on the data of both studies, are that it: does not usually precede the arrival of police in such disputes; is associated with interpersonal closeness of the disputants; is associated with poverty rather than race; and is as likely in a middle-class small city as in a poor inner-city area. Tentative conclusions about the use of alcohol in police-managed interpersonal disputes are: that these disputes are not usually influenced by alcohol use; and that assaultiveness is not related to alcohol use in such disputes.


Effect of Corynebacterium parvum on peripheral blood platelets.
The level of peripheral blood platelets was determined after i.v. injection of Corynebacterium parvum in normal C57BL mice and in those bearing the Lewis lung carcinoma. Twenty minutes after injection of a formalin-killed active strain (CN6134, (CN6134, which inhibited tumour metastases) or a killed inactive strain (CN 5888, which did not inhibit metastases) the number of circulating blood platelets was reduced by 50%. The level of platelets returned to control values by 8 h after the active, and by approximately 3 days after the inactive strain. The active strain alone caused a second and prolonged fall in platelet numbers, from approximately 16 h to 21 days after injection. Heparin given 3 X weekly to these mice restored the platelet count to normal values by 10 days after injection of active-strain C. parvum. The level of platelets in tumour-bearing mice was essentially similar to that in normal mice. Possible causes of the thrombocytopenia and the significance of platelets in metastasis are discussed.


Neonatal Appendicitis and Incarcerated Inguinal Hernia: Case Report and Review of the Literature.
Neonatal appendicitis carries a high mortality rate. We describe a peculiar case presented after an incarcerated hernia manually reduced in a 4-week-old male neonate with ipsilateral undescended testis. Laparoscopy allowed a prompt recognition of an unexpected intra-abdominal life-threatening condition. Pathogenesis, treatment, and outcome are discussed on the light of a comprehensive literature review.


Structural Basis of SARS-CoV-2- and SARS-CoV-Receptor Binding and Small-Molecule Blockers as Potential Therapeutics.
Over the past two decades, deadly coronaviruses, with the most recent being the severe acute respiratory syndrome-related coronavirus-2 (SARS-CoV-2) 2019 pandemic, have majorly challenged public health. The path for virus invasion into humans and other hosts is mediated by host-pathogen interactions, specifically virus-receptor binding. An in-depth understanding of the virus-receptor binding mechanism is a prerequisite for the discovery of vaccines, antibodies, and small-molecule inhibitors that can interrupt this interaction and prevent or cure infection. In this review, we discuss the viral entry mechanism, the known structural aspects of virus-receptor interactions (SARS-CoV-2 S/humanACE2, SARS-CoV S/humanACE2, and MERS-CoV S/humanDPP4), the key protein domains and amino acid residues involved in binding, and the small-molecule inhibitors and other drugs that have (as of June 2020) exhibited therapeutic potential. Specifically, we review the potential clinical utility of two transmembrane serine protease 2 (TMPRSS2)-targeting protease inhibitors, nafamostat mesylate and camostat mesylate, as well as two novel potent fusion inhibitors and the repurposed Ebola drug, remdesivir, which is specific to RNA-dependent RNA polymerase, against human coronaviruses, including SARS-CoV-2.


Levosulpiride-loaded nanostructured lipid carriers for brain delivery with antipsychotic and antidepressant effects.
Herein, we investigate the potential of levosulpiride-loaded nanostructured lipid carriers (LEVO-NLCs) for effective brain delivery with anti-psychotic and antidepressant effects. Micro-emulsion method was used to prepare LEVO-NLCs, followed by its optimization using Design Expert®, investigation of the particles properties and entrapment efficiency (%EE). Moreover, in-vitro release, in-vivo plasma and brain kinetic studies of LEVO-NLCs were executed. Anti-psychotic activity of LEVO-NLCs was accomplished in LPS-induced psychosis mice model. Additionally, expressions of neuro inflammatory mediators, neurodegeneration and neuro-inflammation in brain tissues was investigated. The optimized LEVO-NLCs were rounded shaped nanoparticles (157.2 nm) with suitable zeta potential (-29.6 mV), low PDI (0.395) and high EE (83.67 %). No chemical interactions were found, however, the crystalline drug was changed to amorphous. LEVO-NLCs displayed sustained drug release behavior when compared with drug suspension. Moreover, a meaningfully higher AUC (106,642.27 ± 876.44 ng.h/mL) and Cmax (38,534.72 ± 2344.10 ng/mL) of the LEVO-NLCs in brain was observed as compared to the AUC (15,684.33 ± 1005.49 ng.h/mL) and Cmax (7717.56 ± 871.23 ng/mL) of LEVO-Suspension. Similar profiles of both the formulations were perceived in plasma pharmacokinetic studies. Furthermore, LEVO-NLCs exhibited a meaningfully improved anti-psychotic activity in LPS-induced psychosis mice model with reduced immobility time and enhanced struggling time. Likewise, treatment with LEVO-NLCs showed reduced levels of neuro inflammatory markers (p-NF-κB and COX-2) in LPS-induced mice. Additionally, no neuro-degeneration and neuro-inflammation in brain tissues treated with LEVO-NLCs mice group was detected. These results concluded that NLCs may effectively be used for the brain delivery of various active pharmaceutical agents with enhanced biopharmaceutical performance.


A miniaturized wide-angle 2D cytometer.
We present an optical waveguide based cytometer that is capable of simultaneously collecting the light scattered by cells over a wide range of solid angles. Such comprehensive scattering data are a prerequisite for the microstructural characterization of cells. We use latex beads as cell mimics, and demonstrate the ability of this new cytometer to collect back-scattered light in two dimensions (2D). This cytometer is based on a liquid-core optical waveguide, excited by prism coupling, that also serves as the microfluidic channel. In principle, our use of a hemispherical lens allows the collection of scattered light from 0 to 180 degrees in 2D. The experimentally observed positions of the intensity peaks of the back-scattered light agree well with theoretical prediction of scattering from both 4.0- and 9.6-mum diameter latex beads. The position of the bead, relative to the axes of the hemispherical lens and the microchannel, strongly affects the scattering pattern. We discuss a computational method for determining these offsets. We show that wide-angle 2D light scattering patterns of cell-sized latex beads can be observed in a microfluidic-based optical cytometer that uses leaky waveguide mode excitation. This chip-based system is compatible with emerging chip-based technologies.


Clinical Characteristics and Clinical Course of Body Lateropulsion in 47 Patients with Brainstem Infarctions.
In patients with lower lateral medullary infarction (LMI) located under the vestibular nucleus, proprioceptive impairment due to dorsal spinocerebellar tract (DSCT) is considered a pathological condition for body lateropulsion. In patients with brainstem infarction located at or above the level of the vestibular nucleus, other pathways, such as the crossed vestibulothalamic tract (CVTT), are considered responsible. The clinical course of body lateropulsion between each anatomical level of infarction remains unclear. Further, whether body lateropulsion refers to a static or a dynamic symptom also remains unclear. We examined 47 patients who exhibited body lateropulsion and categorized them into four groups: lower LMI under the vestibular nucleus, LMI at the level of the vestibular nucleus, pontine infarction, and midbrain infarction. The patients' time to acquire static upright standing position and gait in a straight line were statistically analyzed by a log-rank test using the Kaplan-Meier method. Body lateropulsion in the static upright position was less frequent in the lower LMI group than in the other groups. Lower LMI primarily affected body lateropulsion in gait. DSCT damage could affect ipsilateral hip joint or leg coordination, causing body lateropulsion in dynamic situation.


Upconversion FRET quantitation: the role of donor photoexcitation mode and compositional architecture on the decay and intensity based responses.
Lanthanide-doped colloidal nanoparticles capable of photon upconversion (UC) offer long luminescence lifetimes, narrowband absorption and emission spectra, and efficient anti-Stokes emission. These features are highly advantageous for Förster Resonance Energy Transfer (FRET) based detection. Upconverting nanoparticles (UCNPs) as donors may solve the existing problems of molecular FRET systems, such as photobleaching and limitations in quantitative analysis, but these new labels also bring new challenges. Here we have studied the impact of the core-shell compositional architecture of upconverting nanoparticle donors and the mode of photoexcitation on the performance of UC-FRET from UCNPs to Rose Bengal (RB) molecular acceptor. We have quantitatively compared luminescence rise and decay kinetics of Er3+ emission using core-only NaYF4: 20% Yb, 2% Er and core-shell NaYF4: 20% Yb @ NaYF4: 20% Yb, 5% Er donor UCNPs under three photoexcitation schemes: (1) direct short-pulse photoexcitation of Er3+ at 520 nm; indirect photoexcitation of Er3+ through Yb3+ sensitizer with (2) 980 nm short (5-7 ns) or (3) 980 nm long (4 ms) laser pulses. The donor luminescence kinetics and steady-state emission spectra differed between the UCNP architectures and excitation schemes. Aiming for highly sensitive kinetic upconversion FRET-based biomolecular assays, the experimental results underline the complexity of the excitation and energy-migration mechanisms affecting the Er3+ donor responses and suggest ways to optimize the photoexcitation scheme and the architecture of the UCNPs used as luminescent donors.


An evaluation of fast neutron irradiation in the treatment of squamous cell carcinoma in cervical lymph nodes.
Analyses have been made of the response of metastatic cervical lymph nodes following neutron therapy, either as part of a randomized trial or in patients treated electively. In the trial patients, the overall regression and local control rates were similar after photons and neutrons. Mobile nodes, less than 3.0 cm, appeared to respond better to neutron therapy, and node masses greater than 3.0 cm had better control after photon therapy. The differences observed however were not statistically significant. There was a highly significant association between the control of the primary tumor and control of nodal disease. No survival advantage for neutrons was observed in association with apparently better control rates in cervical nodes.


Palladium-catalysed cis- and trans-silaboration of terminal alkynes: complementary access to stereo-defined trisubstituted alkenes.
Palladium-catalysed cis- and trans-silaboration of terminal alkynes has been developed via the addition of (chlorodimethylsilyl)pinacolborane, followed by a one-pot conversion of the chloro group on the silicon atom to an isopropoxy group.


Heart failure in acute ischemic stroke.
To determine the impact of heart failure (HF), with preserved or decreased left ventricular function (LVF), on outcome in patients with acute ischemic stroke (AIS). We studied 503 unselected ischemic stroke patients. Poor outcome was defined as moderate-severe disability or death at 90 days. We analyzed the association between poor outcome and HF with preserved LVF or decreased LVF (systolic HF: ejection fraction lower than 50%). We tested this association adjusted by possible confounders in a logistic regression model. 89 patients (17.7 %) had HF; 49 patients (9.7%) with systolic HF, and 40 (8%) patients with HF and preserved LVF. HF with preserved LVF patients were older [79.4 (7.9) vs. 74.3 (10.4), p = 0.013],more likely to be women [p < 0.001,OR = 8.61, 95% CI (3.3-22.6)], and with lower current smoking habits [p = 0.018, OR = 8.77 (1.1-72.6)] than patients with systolic HF. 151 patients (30 %) had poor outcome. We found an independent association with initial stroke severity, systolic HF (adjusted OR = 3.01), HF with preserved LVF (adjusted OR = 2.52), thrombolytic treatment, statin pre-treatment (as protectors) and poor outcome. Both forms of HF (with or without decreased systolic function) are associated with poor outcome in AIS.


Anemia and Red Blood Cell Transfusion in Aneurysmal Subarachnoid Hemorrhage.
Anemia is very common in aneurysmal subarachnoid hemorrhage (aSAH), with approximately half of the aSAH patient population developing moderate anemia during their hospital stay. The available evidence (both physiologic and clinical) generally supports an association of anemia with unfavorable outcomes. Although aSAH shares a number of common mechanisms of secondary insult with other forms of acute brain injury, aSAH also has specific features that make it unique: an early phase (in which early brain injury predominates) and a delayed phase (in which delayed cerebral ischemia and vasospasm predominate). The effects of both anemia and transfusion are potentially variable between these phases, which may have unique considerations and possibly different risk-benefit profiles. Data on transfusion in this population are almost exclusively limited to observational studies, which suffer from significant heterogeneity and risk of bias. Overall, the results are conflicting, with the balance of the studies suggesting that transfusion is associated with unfavorable outcomes. The transfusion targets that are well established in other critically ill populations should not be automatically applied to patients with aSAH because of the unique disease characteristics of this population and the limited representation of aSAH in the clinical trials that established these targets. There are two upcoming clinical trials evaluating transfusion in aSAH that should help clarify specific transfusion targets. Until then, it is reasonable to base transfusion decisions on the current guidelines and use an individualized approach incorporating physiologic and clinical data when available.


Nucleosome breathing facilitates cooperative binding of pluripotency factors Sox2 and Oct4 to DNA.
Critical lineage commitment events are staged by multiple transcription factors (TFs) binding to their cognate motifs, often positioned at nucleosome-enriched regions of chromatin. The underlying mechanism remains elusive due to difficulty in disentangling the heterogeneity in chromatin states. Using a novel coarse-grained model and molecular dynamics simulations, here we probe the association of Sox2 and Oct4 proteins that show clustered binding at the entry-exit region of a nucleosome. The model captures the conformational heterogeneity of nucleosome breathing dynamics that features repeated wrap-unwrap transitions of a DNA segment from one end of the nucleosome. During the dynamics, DNA forms bulges that diffuse stochastically and may regulate the target search dynamics of a protein by nonspecifically interacting with it. The overall search kinetics of the TF pair follows a "dissociation-compensated-association" mechanism, where Oct4 binding is facilitated by the association of Sox2. The cooperativity stems from a change in entropy caused by an alteration in the nucleosome dynamics upon TF binding. The binding pattern is consistent with a live-cell single-particle tracking experiment, suggesting the mechanism observed for clustered binding of a TF pair, which is a hallmark of cis-regulatory elements, has broader implications in understanding gene regulation in a complex chromatin environment.


The central role of tRNA in genetic code expansion.
The development of orthogonal translation systems (OTSs) for genetic code expansion (GCE) has allowed for the incorporation of a diverse array of non-canonical amino acids (ncAA) into proteins. Transfer RNA, the central molecule in the translation of the genetic message into proteins, plays a significant role in the efficiency of ncAA incorporation. Here we review the biochemical basis of OTSs for genetic code expansion. We focus on the role of tRNA and discuss strategies used to engineer tRNA for the improvement of ncAA incorporation into proteins. The engineering of orthogonal tRNAs for GCE has significantly improved the incorporation of ncAAs. However, there are numerous unintended consequences of orthogonal tRNA engineering that cannot be predicted ab initio. Genetic code expansion has allowed for the incorporation of a great diversity of ncAAs and novel chemistries into proteins, making significant contributions to our understanding of biological molecules and interactions. This article is part of a Special Issue entitled "Biochemistry of Synthetic Biology - Recent Developments" Guest Editor: Dr. Ilka Heinemann and Dr. Patrick O'Donoghue.


Single-step synthesis of gold-silver alloy nanoparticles in ionic liquids by a sputter deposition technique.
The simultaneous sputter deposition of gold and silver onto ionic liquids formed bimetallic alloy nanoparticles, which exhibited composition-sensitive surface plasmon resonance, the peak wavelength being red-shifted with an increase in the surface area of the gold foil targets sputtered.


Systematics and evolution of the genus Torrubiella (Hypocreales, Ascomycota).
Torrubiella is a genus of arthropod-pathogenic fungi that primarily attacks spiders and scale insects. Based on the morphology of the perithecia, asci, and ascospores, it is classified in Clavicipitaceae s. lat. (Hypocreales), and is considered a close relative of Cordyceps s. 1., which was recently reclassified into three families (Clavicipitaceae s. str., Cordycipitaceae, Ophiocordycipitaceae) and four genera (Cordyceps s. str, Elaphocordyceps, Metacordyceps, and Ophiocordyceps). Torrubiella is distinguished morphologically from Cordyceps s. lat. mainly by the production of superficial perithecia and the absence of a well-developed stipitate stroma. To test and refine evolutionary hypotheses regarding the placement of Torrubiella and its relationship to Cordyceps s. lat., a multi-gene phylogeny was constructed by conducting ML and Bayesian analyses. The monophyly of Torrubiella was rejected by these analyses with species of the genus present in Clavicipitaceae, Cordycipitaceae, and Ophiocordycipitaceae, and often intermixed among species of Cordyceps s. lat. The morphological characters traditionally used to define the genus are, therefore, not phylogenetically informative, with the stipitate stromata being gained and/or lost several times among clavicipitaceous fungi. Two new genera (Conoideocrella, Orbiocrella) are proposed to accommodate two separate lineages of torrubielloid fungi in the Clavicipitaceae s. str. In addition, one species is reclassified in Cordyceps s. str. and three are reclassified in Ophiocordyceps. The phylogenetic importance of anamorphic genera, host affiliation, and stipitate stromata is discussed.


Recent advances in laboratory tests for autoantibodies to thyrotropin receptor protein in Graves' disease.
Graves' disease is characterized by the presence of autoantibodies with variable reactivities to TSH receptor protein. These autoantibodies are heterogeneous in nature and may act as TSH agonists or infrequently as TSH antagonists (blocking antibodies). With new technical advances, it has become possible to quantitate these antibodies and their reactivities in clinical laboratories. Technical aspects of these assays and their clinical utility in the diagnosis and management of patients with Graves' disease are discussed.


Using children's picture books for reflective learning in nurse education.
One way in which nursing students may build their practice is through reflective learning from stories. Stories in children's literature offer a special source of narratives that enable students to build empathy and to examine and reconstruct their personal concepts around human experience. Illustrated storybooks written for children are a particularly attractive teaching resource, as they tend to be short, interesting, colourful and easy to read. Yet, little has been written about using such books as a reflective learning tool for nursing students. In this article we describe how we use two children's books and McDrury and Alterio's (2002) 'Reflective Learning through Storytelling' model to educate first year nursing students about loss, grief and death.


Cervical cancer and cytology screening in New Zealand.
Cytology screening, used in New Zealand since 1955 at an intensity comparable to that in Canada generally, has not favourably affected incidence and mortality rates for cervical cancer; these have actually risen significantly in 20 to 34-year-old New Zealand women. Canadian claims that mortality falls are related to intensity of cytological screening are not justifiable, so that the significance of the 'pre-cancers' revealed by cytology and the value of population screening would seem to be doubtful.


Assessing the utility of universal preprocedure severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) polymerase chain reaction (PCR) testing at a cancer center.
We implemented preprocedure severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing for cancer patients. The overall positivity rate was 0.3%. Preprocedure testing may be limited to operative procedures as community rates of SARS-CoV-2 exceed 7%. Nonoperative aerosol-generating procedures may be performed without preprocedure testing, wearing the appropriate personal protective equipment.


Entamoeba moshkovskii infections in Sydney, Australia.
Entamoeba moshkovskii is considered to be a free-living ameba, which is morphologically similar, but biochemically and genetically different, to Entamoeba histolytica and Entamoeba dispar. However, recent studies have suggested that E. moshkovskii may be a "potential" pathogen, with infections giving rise to diarrhea and other intestinal disorders. Microscopy and polymerase chain reaction (PCR) targeting the 18S ribosomal (r) DNA was performed on fecal samples collected from patients presenting with diarrhea and other gastrointestinal disorders (test group), as well as fecal samples collected from healthy controls (control group). Of the 110 patients microscopically positive for the Entamoeba species, 55/110 (50%) samples were positive for E. moshkovskii in the test group of patients presenting with diarrhea. E. moshkovskii was the only pathogen detected (including bacteria or viruses) in 3/55 (5.5%) of the test group of patients presenting with diarrhea and abdominal pain. The DNA of E. moshkovskii was not detected in the fecal samples collected from the healthy controls. These results suggest that E. moshkovskii may not simply be a commensal of the human gastrointestinal tract and provides evidence for E. moshkovskii as a "potential" pathogen in the case of diarrhea and other gastrointestinal disorders. Further studies are needed to determine the role of E. moshkovskii in causing diarrheal diseases in our population.