Isolation and characterization of Arcobacter spp. from fresh seafood and the aquatic environment.
Arcobacter is an emerging pathogen associated with foods of animal origin. Members of the genus Arcobacter are increasingly being isolated from fish, shellfish and the aquatic environment. In the present study, we analyzed fish, shellfish and water samples for the presence of Arcobacter spp. by conventional isolation as well as by direct PCR on the enrichment broth. Of 100 samples comprising of 42 finfish, 34 shellfish and 24 water samples analyzed, Arcobacter spp. was isolated from 8 (19%) finfish, 5 (14.7%) shellfish and 5 (20.8%) water samples. Arcobacter DNA was detected in 24 (24%) samples by direct PCR on the enrichment broth. Based on m-PCR specific to different Arcobacter spp. and 16S rRNA sequence analyses, majority (19) of the isolates were identified as Arcobacter butzleri, while two isolates were Arcobacter mytili. All Arcobacter butzleri isolates harbored putative virulence genes cadF, ciaB, mviN, pldA, tlyA and cj1349. The two isolates of A. mytili harbored mviN and cj1349 genes only. The study highlights emerging problem of the contamination of aquatic environment and fresh seafood with potentially pathogenic Arcobacter spp.


Genetic analysis of human immunodeficiency virus type 1 nef in Portugal: subtyping, identification of mosaic genes, and amino acid sequence variability.
Extending our previous genetic characterization of human immunodeficiency virus type 1 (HIV-1) strains circulating in Portugal, we here report the first phylogenetic and putative amino acid sequence variability analyses of nef accessory gene. Viral sequences (n = 53) were amplified by nested PCR from proviral DNA purified from peripheral blood mononuclear cells of HIV-1 infected individuals (n = 49). Phylogenetic inference analysis demonstrated a distribution of the viral sequences between subtypes A (sub-subtype A1), B, D, F (sub-subtype F1), G, H, and J, with subtypes G and B accounting altogether for more than half of the genotypes found. A significant number of the proviral DNA sequences analyzed (18.4%) were shown to correspond to intragenic nef recombinants, with the majority having the typical CRF02_AG nef structure. In addition, three novel intragenic recombinant structures were found (B/G/B, CRF02_AG/H, and D/G). From phylogenetic analysis, it was concluded that part of the non-recombinant nef genes might have actually been amplified from mosaic viruses: CRF06_cpx, CRF14_BG, and a new envA/nefJ recombinant. While comparing all the putative Nef sequences, significant amino acid sequence variability was observed. However, most of the described nef functional motifs were relatively well conserved in the majority of the sequences analyzed and numerous amino acid changes fell outside these regions. The results presented unambiguously endorse the high level of complexity of HIV-1 epidemics in Portugal.


Enteroendocrine peptides, growth, and the microbiome during the porcine weaning transition.
Growth rate in pigs can be affected by numerous factors that also affect feeding behavior and the microbiome. Recent studies report some communication between the microbiome and the enteroendocrine system. The present study examined if changes in the piglet microbiome between birth and during the weaning transition can be correlated either positively or negatively with growth rate and plasma concentrations of enteroendocrine peptides. During the post-weaning transition, a 49% reduction in average daily gain was observed at day 24 (P < 0.05) relative to day 21. Pigs recovered by day 28 with body weight and average daily gain increases of 17% and 175%, respectively relative to day 24 and the highest rate of gain was measured at day 35 (462 g/day). The time interval between day 21-24 had the highest number of correlations (n = 25) between the relative abundance differences in taxa over time and corresponding percent weight gain. Amplicon sequence variants with the greatest correlation with percent weight gain between day 21-24 belonged to families Prevotellaceae NK3B31 (ρ = 0.65, P < 0.001), Veillonellaceae (ρ = 0.63, P < 0.001) and Rikenellaceae RC9 (ρ = 0.62, P < 0.001). Seven taxa were positively correlated with percent weight gain between day 24-28. Eight taxa were positively correlated with percent weight gain between day 28-35, of which four were Clostridia. Only Lactobacillus reuteri was positively correlated across both day 24-28 and day 28-35 analyses. Insulin-like growth factor 1 (IGF-1; R2 = 0.61, P < 0.001), glucose-dependent insulinotropic polypeptide (GIP; R2 = 0.20, P < 0.001), glucagon-like peptide 1 (GLP-1; R2 = 0.51, P < 0.001), and glucagon-like peptide 2 (GLP-2; R2 = 0.21, P < 0.001) were significantly associated with the piglet fecal community NMDS, while serotonin showed no significant association (R2 = 0.03, P = 0.15). Higher concentrations of GLP-1 and GLP-2 characterized day 1 fecal communities, while GIP levels had the strongest relationship primarily with samples ordinated with the day 21 cluster. Demonstration of an association of certain taxa with individual gut peptides at specific ages suggests the potential for the microbiome to elicit changes in the gut enteroendocrine system during early postnatal development in the pig.


The Impact of Early-Life Exposure to Antimicrobials on Asthma and Eczema Risk in Children.
We examined recent research on associations of prenatal and early-childhood exposure to the antimicrobial compounds, triclosan, and parabens, with the risk of asthma and eczema in children. We will discuss potential biological mechanisms of this association and highlight strengths and limitations of the study design and exposure assessment of current findings. Results of available toxicological and epidemiologic studies indicate a potential link of triclosan and paraben exposures with asthma and eczema in children, as well as changes in microbiome diversity and immune dysfunction, which could possibly mediate an association with the health outcomes. A small number of studies suggest that triclosan and paraben exposures could be related to the risk of asthma and eczema in children. Although current findings are far from conclusive, there is emerging evidence that changes in microbiome diversity and immune function from antimicrobial exposure may mediate these relations.


Brief ganglioside treatment produces delayed enhancement of functional recovery after medial septal lesions.
The effects of a 5-day ganglioside (GM1) treatment (30 mg/kg) on body weight and water intake subsequent to medial septal lesions were evaluated for 44 consecutive days. In addition, activity, rearing, and repetitive motor acts were measured on postsurgery days 5, 10, 40, and 60. The rate of increase in the body weights of rats with medial septal lesions treated with GM1 was equivalent to that of controls, while untreated rats with such lesions had reduced body weights. Rats with medial septal lesions treated with GM1 also exhibited movement times and frequency of repetitious motor acts similar to those of control rats by postsurgery day 60. No differences were found in water intake between any of the groups. Rats with medial septal lesions, whether treated with GM1 or not, had equivalent frequencies of rearings that were lower than control rats. This study emphasizes that even brief regimes of GM1 administration can exert behavioral changes in brain-damaged rats well after the treatment was administered, i.e. 40-60 days after surgery.


Oral PowderJect: a novel system for administering local anaesthetic to the oral mucosa.
To assess the feasibility of using an Oral PowderJect (OPJ) to safely deliver a dose of dry powdered anaesthetic to the oral mucosa, producing an analgesic effect. Single centre: Part 1. An open, non-randomised safety study to check for mucosal damage; Part 2. A double blind sham controlled study to test the anaesthetic effect. General practice. Adult, healthy volunteers (4 male, 10 female). Part 1. An OPJ was used to deliver powdered lidocaine hydrochloride to the mucosal surface which was then checked visually for damage. Part 2. An OPJ containing lidocaine hydrochloride (active) or an empty OPJ (sham) was fired at the oral mucosa. The treated area and an untreated (control) site were probed with the back end of a dental needle. The OPJ delivery caused no visible mucosal damage. The median VAS score for pain on blunt probing was 10 for the OPJ active sites. This was significantly lower than the median VAS score for the sham sites at 30 (P = 0.0033) and the control sites at 58 (P < 0.0001). The OPJ can safely deliver powdered lidocaine hydrochloride to the oral mucosa without causing tissue damage. The OPJ delivery of powdered lidocaine hydrochloride can significantly reduce the pain from a blunt needle probe at 1 minute post delivery.


Dissection of the contribution of individual domains to the ATPase mechanism of Hsp90.
Hsp90 is a dimeric, ATP-regulated molecular chaperone. Its ATPase cycle involves the N-terminal ATP binding domain (amino acids (aa) 1-272) and, in addition, to some extent the middle domain (aa 273-528) and the C-terminal dimerization domain (aa 529-709). To analyze the contribution of the different domains and the oligomeric state on the progression of the ATPase cycle of yeast Hsp90, we created deletion constructs lacking either the C-terminal or both the C-terminal and the middle domain. To test the effect of dimerization on the ATPase activity of the different constructs, we introduced a Cys residue at the C-terminal ends of the constructs, which allowed covalent dimerization. We show that all monomeric constructs tested exhibit reduced ATPase activity and a decreased affinity for ATP in comparison with wild type Hsp90. The covalently linked dimers lacking only the C-terminal domain hydrolyze ATP as efficiently as the wild type protein. Furthermore, this construct is able to trap the ATP molecule similar to the full-length protein. This demonstrates that in the ATPase cycle, the C-terminal domain can be replaced by a cystine bridge. In contrast, the ATPase activity of the artificially linked N-terminal domains remains very low and bound ATP is not trapped. Taken together, we show that both the dimerization of the N-terminal domains and the association of the N-terminal with the middle domain are important for the efficiency of the ATPase cycle. These reactions are synergistic and require Hsp90 to be in the dimeric state.


Effects of the virulence plasmid ColV, I-K94 on the sensitivity of Escherichia coli to putative environmental inhibitory agents.
Derivatives of Escherichia coli carrying the virulence plasmid, ColV, I-K94 were more resistant than the ColV- parents to phage Mel but were more sensitive to the hydrophobic inhibitors deoxycholate, erythromycin and lysozyme. The basis for these changes in sensitivity has been examined in ColV+ mutants with altered colicin or VmpA protein levels and in ColV+ strains with repressed transfer properties.


Prophylactic placement of external iliac artery balloon catheter in a patient with intrapelvic prosthesis migration after hemiarthroplasty: A case report.
Intrapelvic prosthesis migration is a rare but serious complication of bipolar hemiarthroplasty in femoral neck fractures. The external iliac artery is one of the most frequently damaged arteries during the removal of a migrated implant from the pelvic region. This report describes a case in which prophylactic placement of an external iliac artery balloon catheter was performed to reduce blood loss in the event of vascular injury during implant removal surgery in the pelvic region.


Adolescent depression in Japan: as studied from Shimoda's theory and Harter's theory.
Through the daily treatment of depressive adolescents we have come to feel that such cases present with a common specific state at onset that includes hypersensitivity and a tendency to neuroticism. This state is reminiscent of the introverted character of children reported by Shimoda in 1929 and of immodithymic character that he later proposed as the pre-pathological character of bipolar disorders. In contrast, we supposed that it might be important to evaluate these adolescents from the low self-worth theory and to compare adolescent psychopathology between the USA and Japan. In order to ascertain these issues, we performed a clinical survey of the junior and high school students who came to outpatient clinics, and thereby demonstrated the necessity of understanding the depressive adolescents from the standpoint of character theory and self-worth theory. We then attempted some field work with ordinary junior high school children as object using Harter's self-perception profile and Kovac's CDI. The results showed that depressive tendencies and self-worth differed greatly among Japanese junior high school children and American counterparts. We have used these findings to discuss the psychopathology of depressive tendencies in Japanese adolescents.


Longitudinal Analysis of Microbiota in Microalga Nannochloropsis salina Cultures.
Large-scale open microalgae cultivation has tremendous potential to make a significant contribution to replacing petroleum-based fuels with biofuels. Open algal cultures are unavoidably inhabited with a diversity of microbes that live on, influence, and shape the fate of these ecosystems. However, there is little understanding of the resilience and stability of the microbial communities in engineered semicontinuous algal systems. To evaluate the dynamics and resilience of the microbial communities in microalgae biofuel cultures, we conducted a longitudinal study on open systems to compare the temporal profiles of the microbiota from two multigenerational algal cohorts, which include one seeded with the microbiota from an in-house culture and the other exogenously seeded with a natural-occurring consortia of bacterial species harvested from the Pacific Ocean. From these month-long, semicontinuous open microalga Nannochloropsis salina cultures, we sequenced a time-series of 46 samples, yielding 8804 operational taxonomic units derived from 9,160,076 high-quality partial 16S rRNA sequences. We provide quantitative evidence that clearly illustrates the development of microbial community is associated with microbiota ancestry. In addition, N. salina growth phases were linked with distinct changes in microbial phylotypes. Alteromonadeles dominated the community in the N. salina exponential phase whereas Alphaproteobacteria and Flavobacteriia were more prevalent in the stationary phase. We also demonstrate that the N. salina-associated microbial community in open cultures is diverse, resilient, and dynamic in response to environmental perturbations. This knowledge has general implications for developing and testing design principles of cultivated algal systems.


Identification and quantification of anthocyanins, flavonoids, and phenolic acids in flowers of Rhododendron arboreum and evaluation of their antioxidant potential.
The current study aimed to investigate the anthocyanins, non-anthocyanins (flavonoids and phenolic acids), and free radicals scavenging potential in the flowers of Rhododendron arboreum using ultra high performance liquid chromatography with ion mobility quadrupole time of flight tandem mass spectrometry. A total of 25 constituents including nine anthocyanins, six phenolic acids, and ten flavonoids were identified in the flower extract. The major anthocyanins identified were cyanidin-3-O-β-galactoside (1), cyanidin-3-O-α-arabinoside (4), and cyanidin-3-O-rhamnoside (8), while quercetin glycosides were the main identified flavonoids in R. arboreum flowers. Additionally, ultra high performance liquid chromatography methods were developed and validated for the quantification of nine compounds (anthocyanins, flavonoid glycosides, and phenolic acids); five of them were quantified using internal standards. The extracts were analyzed for total phenolics (123.6 mg GAE/g), anthocyanin content (1.76% w/w), and evaluated for antioxidant properties against 2,2-diphenyl-1-picrylhydrazyl radical (IC50 : 102.06 and 96.92 μg/mL) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical cation (112.25 and 45.59 μM TE/g) assays. The profiling of R. arboreum for anthocyanins is reported for the first time. The findings suggest that the flowers are a promising source of bioactive constituents and could be used as functional food, antioxidants, and nutraceuticals.


Inhibition of class I HDACs attenuates renal interstitial fibrosis in a murine model.
Renal interstitial fibrosis is the most common of all the forms of chronic kidney disease (CKD). Research has shown that histone deacetylases (HDACs) participate in the process leading to renal fibrosis. However, the effects of class I HDAC inhibitors on the mechanisms of onset and progression of renal interstitial fibrosis are still unclear. Here, we present the effects and mechanisms of action of FK228 (a selective inhibitor of class I HDACs) in the murine model of unilateral ureteral obstruction (UUO) and in vitro models. We investigated the antifibrotic role of FK228 in a murine model of UUO. We used two key effector cell populations, rat renal interstitial fibroblasts and renal tubular epithelial cells exposed to recombinant transforming growth factor-beta 1 (TGF-β1), to explore the mechanistic pathways among in vitro models. The results indicated that FK228 significantly suppressed the production of extracellular matrix (ECM) in both in vivo and in vitro models. FK228 inhibited renal fibroblast activation and proliferation and increased the acetylation of histone H3. We found that FK228 also inhibited the small mothers against decapentaplegic (Smad) and non-Smad signaling pathways. So FK228 could significantly suppress renal interstitial fibrosis via Smad and non-Smad pathways. FK228 may be the basis for a new and effective medicine for alleviating renal fibrosis in the future.


Changes in normalized mutual information in response to strength training: An ancillary analysis of a quasi-randomized controlled trial.
The aim of the present investigation was twofold. (1) to assess test-retest reliability of normalized mutual information (NMI) values extracted from the surface electromyography (sEMG) signal of muscles pairs of the upper body during dynamic bench press at a high load, and (2) to assess changes in NMI values from before to after a five-week quasi-randomized controlled bench press training intervention. For test-retest reliability, 20 strength trained males (age 25 ± 2 years, height 1.81 ± 0.07 m) performed two three-repetition maximum (3RM) tests in bench press, while sEMG was recorded from six upper body muscles. Tests were separated by 8.2 ± 2.9 days. For the training intervention, 17 male participants (age 26 ± 5 years, height 1.80 ± 0.07 m) trained bench press specific strength training for 5 weeks (TRA), while 13 male participants (age 23 ± 3 years, height 1.80 ± 0.08 m) constituted a control group (CON). 3RM bench press test and sEMG recordings were carried out before and after the intervention period. The NMI values ranged from poor to almost perfect reliability, with the majority displaying substantial reliability. TRA displayed a significant decrease in NMI values during the concentric phase for two agonist-agonist muscle pairs, while one agonist-agonist and two agonist-antagonist muscle pairs increased the NMI values during the eccentric phase. The observed changes did not exceed the minimal detectable threshold, and we therefore cannot surely ascertain that the changes observed in NMI values reflect genuine neural adaptations.


Palmitate and Nitric Oxide Regulate the Expression of Spexin and Galanin Receptors 2 and 3 in Hypothalamic Neurons.
Spexin (SPX) is a novel satiety factor that putatively binds the galanin receptors R2 and R3 (GalR2/R3). SPX reduces body weight, and circulating SPX is decreased in obesity. It is unknown how SPX and its receptors are regulated in the hypothalamus, critical for energy homeostasis. We therefore examined the regulation of hypothalamic Spx, GalR2 and GalR3 gene expression in mouse primary and immortalized hypothalamic neurons. We report that Spx, GalR2 and GalR3 mRNA levels were regulated by acute treatments of palmitate, a dietary saturated fatty acid, as well as the nitric oxide (NO) donor sodium nitroprusside (SNP), but through a pathway independent of cyclic GMP and protein kinase G. Additionally, the palmitate- and NO-mediated induction of Spx and galanin receptors was blocked with the PKC inhibitor k252c. Furthermore, palmitate induced mRNA levels of endoplasmic reticulum (ER) stress markers, including Chop, Grp78 and Bax/Bcl2, as well as C/ebp-β, whereas SNP induced Bax/Bcl2 and C/ebp-β. Transcriptional changes in Spx, GalR2, GalR3, C/ebp-β and ER stress marker mRNAs were blocked by pre-treatment with at least one of the chemical chaperones PBA or TUDCA. We also describe the presence of OCT-1 and C/EBP-β response elements in the 5' regulatory region of Spx and demonstrate that SNP increases binding of C/EBP-β to this region, but not Oct-1 mRNA nor OCT-1 binding. Our findings suggest an acute modulation of anorexigenic SPX signaling by palmitate and NO. Furthermore, ER stress and C/EBP-β appear to mediate the changes in Spx, GalR2 and GalR3 in hypothalamic neurons.


Emotional reaction to fragile X premutation carrier tests among infertile women.
Female fragile X premutation carriers are at approximately 10-fold increased risk of premature ovarian failure (follicle stimulating hormone >40 mIU/mL, amenorrhea, age <40). A milder degree of premature ovarian aging (diminished ovarian reserve, where follicle stimulating hormone levels are typically 10-20 mIU/mL) results in infertility. Approximately 10% of fertility clinic patients have this diagnosis. A cohort of 20 women diagnosed with diminished ovarian reserve provided a blood specimen (confidential results), and completed structured questionnaires that assessed emotional reactions to potentially being a premutation carrier (pretest questionnaire, n = 20) and the posttest known carrier status (3 month follow-up questionnaire, n = 18 non-carriers). Responses were measured using 9-point scales, and analyzed with Fisher exact and Wilcoxon exact tests. While most participants did not view fragile X premutations as a serious medical condition, perceptions of seriousness were positively correlated with anger and regret about not knowing sooner of the potential association of these premutations with infertility. Overall, when women (pretest) imagined themselves as carriers, their self-esteem and Health Orientation Scale responses were unchanged with the exception of feeling more afraid (p = 0.004). Despite strongly wishing for negative test results, they were glad to know there might be a medical explanation for their infertility.


Substance P antagonist inhibits vasomotor responses elicited from ventral medulla in rat.
Application of kainic acid to the ventral medulla in rats leads to increases in blood pressure and heart rate. These responses can be attenuated or reversed by infusion of the substance P antagonist--(D-Pro2,D-Trp7,9)-substance P into the spinal subarachnoid space. Kainic acid elicits normal pressor responses from rats that have been treated intracisternally with the serotonin neurotoxin 5,7-dihydroxytryptamine. These results suggest that the substance P-like neurons of the ventral medulla may play a role in maintaining vasomotor tone.


Technique for creating the anterior cruciate ligament femoral socket: optimizing femoral footprint anatomic restoration using outside-in drilling.
The purpose of this study was to investigate and optimize anterior cruciate ligament (ACL) femoral outside-in drilling technique with a goal of anatomic restoration of the footprint morphologic length, width, area, and angular orientation. Ex vivo, computer navigation was used to create virtual 3-dimensional maps of femoral bone tunnels for ACL drill guide pin insertion paths on small, medium, and large models of averaged femora considering various pin insertion angles to the femur. We then determined which pin insertion angle resulted in an ACL femoral footprint optimally matching normal human anatomic length, width, area, and angular orientation of the footprint long axis. During outside-in drilling of the ACL femoral socket, a guide pin entrance angle of 60° to a line perpendicular to the femoral anatomic axis, combined with a guide pin entrance angle of 20° to the transepicondylar axis, results in the closest approximation of the gold standard of normal anatomic morphology of the human knee ACL femoral footprint length, width, area, and angular orientation. During outside-in drilling of the ACL femoral socket, a guide pin entrance angle of 60° to a line perpendicular to the femoral anatomic axis, combined with a guide pin entrance angle of 20° to the transepicondylar axis, results in optimal reconstruction of the normal human anatomic ACL femoral footprint length, width, area, and angular orientation. We describe arthroscopic landmarks for anatomic ACL femoral socket creation that may be considered by practicing arthroscopic surgeons in the operating room, without open dissection or fluoroscopy and unaffected by type of drill guide or variations in the thickness of the femoral soft-tissue envelope.


Pathogenesis of atherosclerosis: a possible relation to infection.
Atherosclerosis is the main underlying cause of coronary heart disease, which in turn is the most common cause of death in the industrialized world. An acute event in coronary heart disease is typically precipitated by thrombosis occurring at the site of atherosclerotic plaque disruption. Atherosclerotic plaques consist of a fibrous cap overlying a lipid-rich core. Many cell types are involved in their formation, including platelets, endothelial cells, activated monocytes, macrophages derived from monocytes and smooth muscle cells. The currently accepted hypothesis is that atherosclerosis develops as a response to injury and that it is primarily a chronic inflammatory condition. The endothelium plays an important role in regulating vascular blood flow and it is now apparent that endothelial dysfunction is an important contributor to the pathogenesis of atherosclerosis. There is growing evidence that infection may be a risk factor for atherosclerosis and myocardial infarction. Numerous studies have reported associations between human coronary heart disease (CHD) and bacterial and viral infections. At present, interest is focused on the potential aetiological role of C. pneumoniae which has been repeatedly identified, using various diagnostic techniques, in atherosclerotic lesions. There is also increasing seroepidemiological evidence of the association between C. pneumoniae and CHD. The role of this organism in atherosclerosis may be analogous to that of chronic C. trachomatis infection in trachoma.


Structure-based design of cyclooxygenase-2 selectivity into ketoprofen.
We have recently described how to achieve COX-2 selectivity from the non-selective inhibitor indomethacin (1) using a combination of a pharmacophore and computer 3-D models based on the known X-ray crystal structures of cyclooxygenases. In the present study we have focused on the design of COX-2 selective analogues of the NSAID ketoprofen (2). The design is similarly based on the combined use of the previous pharmacophore together with traditional medicinal chemistry techniques motivated by the comparative modeling of the 3-D structures of 2 docked into the COX active sites. The analysis includes use of the program GRID to detect isoenzyme differences near the active site region and is aimed at suggesting modifications of the basic benzophenone frame of the lead compound 2. The resulting series of compounds bearing this central framework is exemplified by the potent and selective COX-2 inhibitor 17 (LM-1669).


The distribution of antibodies to Border disease virus among sheep in England and Wales.
Within a sample of the sheep population of England and Wales, 10.8 per cent of 3506 individuals had immunodiffusion test antibody to bovine viral diarrhoea mucosal disease virus antigen. There were marked differences between various geographical regions in the proportion of sheep with antibody, and Border disease may be more common in some areas than disease records indicate.


Carbon and oxygen isotope characteristics of carbonate rocks in the Mesoproterozoic Jixian System of the Ordos Basin and their implications.
The paleoenvironment of Jixian carbonate rocks in the Mesoproterozoic Ordos Basin is studied by carbon and oxygen isotope analyses, diagenetic environment analysis, and the restoration of paleosalinity and paleotemperature. The results indicate that the carbonate rocks of the Jixian System have always been in a near-surface environment and have not been deeply buried. The ranges of variation in δ13CPDB and δ18OPDB are relatively narrow, ranging from - 5.75 to 1.41‰ and - 8.88 to - 4.01‰, respectively, which is consistent with the stable tidal flat sedimentary environment during the Mesoproterozoic in the study area. The paleosalinity (Z) values range from 111.7 to 127.1, and the paleotemperature (T) values range from 32.7 to 57.33 °C, indicating a relatively warm paleoclimatic environment during the Mesoproterozoic era in the study area. The analysis shows that in a warm paleoclimatic environment, although carbon and oxygen isotopes, Z, and T have certain fluctuations, their ranges are relatively small, reflecting to some extent the stable tectonic environment of the study area during the Mesoproterozoic era. Comprehensive research shows that the Ordos Basin had a warm climate and a stable tectonic environment in the Mesoproterozoic, which may be a good response to the North China Block's position near the equator and continuous thermal subsidence in the Mesoproterozoic.


Melanoma cell lysosome secretory burst neutralizes the CTL-mediated cytotoxicity at the lytic synapse.
Human melanoma cells express various tumour antigens that are recognized by CD8(+) cytotoxic T lymphocytes (CTLs) and elicit tumour-specific responses in vivo. However, natural and therapeutically enhanced CTL responses in melanoma patients are of limited efficacy. The mechanisms underlying CTL effector phase failure when facing melanomas are still largely elusive. Here we show that, on conjugation with CTL, human melanoma cells undergo an active late endosome/lysosome trafficking, which is intensified at the lytic synapse and is paralleled by cathepsin-mediated perforin degradation and deficient granzyme B penetration. Abortion of SNAP-23-dependent lysosomal trafficking, pH perturbation or impairment of lysosomal proteolytic activity restores susceptibility to CTL attack. Inside the arsenal of melanoma cell strategies to escape immune surveillance, we identify a self-defence mechanism based on exacerbated lysosome secretion and perforin degradation at the lytic synapse. Interfering with this synaptic self-defence mechanism might be useful in potentiating CTL-mediated therapies in melanoma patients.


Amphibian deformities and Ribeiroia infection: an emerging helminthiasis.
Since their widespread appearance in the mid-1990s, malformed amphibians have evoked fear, as well as fascination within the scientific and public communities. Recent evidence from field and laboratory studies has implicated infection by a digenetic trematode--Ribeiroia ondatrae--as an important cause of such deformities. Ribeiroia spp. have a complex life cycle involving planorbid snails, amphibians and water birds. Under natural conditions, malformations might promote parasite transmission by increasing the susceptibility of infected amphibians to predation by definitive hosts. However, with respect to the recent outbreak of deformities, we suggest that exogenous agents (e.g. pesticides, nutrient run-off, introduced fishes) might be interacting with Ribeiroia, resulting in elevated infection levels, and we highlight the need for studies incorporating multiple stressor dynamics to further explore this problem.


Molecular MRI differentiation between primary central nervous system lymphomas and high-grade gliomas using endogenous protein-based amide proton transfer MR imaging at 3 Tesla.
To show the ability of using the amide proton transfer-weighted (APTW) MRI signals as imaging biomarkers to differentiate primary central nervous system lymphomas (PCNSLs) from high-grade gliomas (HGGs). Eleven patients with lymphomas and 21 patients with HGGs were examined. Magnetization-transfer (MT) spectra over an offset range of ± 6 ppm and the conventional MT ratio (MTR) at 15.6 ppm were acquired. The APTW signals, total chemical-exchange-saturation-transfer signal (integral between 0 and 5 ppm, CEST total), and MTR signal were obtained and compared between PCNSLs and HGGs. The diagnostic performance was assessed with the receiver operating characteristic (ROC) curve analysis. The PCNSLs usually showed more homogeneous APTW hyperintensity (spatially compared to normal brain tissue) than the HGGs. The APTW max, APTW max-min and CEST total signal intensities were significantly lower (P < 0.05, 0.001 and 0.05, respectively), while the APTW min and MTR were significantly higher (both P < 0.01) in PCNSL lesions than in HGG lesions. The APTW values in peritumoral oedema were significantly lower for PCNSLs than for HGGs (P < 0.01). APTW max-min had the highest area under the ROC curve (0.963) and accuracy (94.1 %) in differentiating PCNSLs from HGGs. The protein-based APTW signal would be a valuable MRI biomarker by which to identify PCNSLs and HGGs presurgically. PCNSLs overall showed more homogeneous APTW hyperintensity than HGGs. Maximum APTW signals were lower in PCNSL lesions than in HGG lesions. MTR signals were higher in PCNSLs than in HGGs. APTW heterogeneity had the highest accuracy in differentiating PCNSLs from HGGs.


Early effects of abrupt reduction of local pressure on the forearm and its circulation.
1. The pressure at the surface of a segment of forearm enclosed in a plethysmograph was abruptly reduced from atmospheric to -20 to -120 mm Hg.2. Forearm circumference (equivalent to the volume of a small segment of forearm (V(f))) was measured with a strain gauge. Pressure was measured in the plethysmograph (P(p)), in veins exposed (P(ve)) and not exposed (P(vne)) to suction, in the brachial artery not exposed to suction (P(bane)) and in forearm tissue (P(t)).3. Reduction of P(p) caused increase of V(f). This was not due to gas evolution, since bubbles would not be liberated at the pressures employed. Nor was increase of V(f) due to venous backflow since P(ve) fell, but P(vne) did not, even with upper arm circulation occluded or when P(vne) was raised by venous occlusion prior to reduction of P(p).4. Reduction of P(p) temporarily arrested venous outflow since P(ve) < P(vne) < P(bane) for 30 sec. With reduction of P(p) 30 sec after occlusion of the upper-arm circulation, P(ve) < P(vne) for > 1 min, indicating that arterial inflow was then minimal.5. Increase of V(f), following reduction of P(p), was therefore due to inflow of arterial blood, of soft tissue or interstitial fluid. Interstitial fluid could flow from regions external to the plethysmograph, or enter as the result of filtration across capillaries. Occlusion of the upper arm circulation was not expected to interfere with motion of forearm soft tissue or the intratissue flow of interstitial fluid. It appears that capillary filtration is small compared with observed blood flow. Therefore subtraction of V(foccl) measured at intervals after reduction of P(p) (upper arm circulation occluded) from V(f) similarly obtained (but upper arm circulation free) appeared to give change of forearm volume due to inflow of arterial blood (DeltaV(b)). V(b), the volume inflow rate of arterial blood during suction, was then obtained.6. Resting forearm flow was 1.8 ml./min/100 ml. in seven normal subjects (average mean arterial blood pressure 86 mm Hg). With P(p) = -90 mm Hg, V(b) was 10.2 ml./min/100 ml. Suction therefore reduced vascular resistance, measured as (P(bane)-P(ve)) /V(b).


Cancer progression and monocyte inflammatory dysfunction: relationship to tumour excision and metastasis.
In a longitudinal study performed before and after surgical resection of a chemically-induced, transplanted rat tumour we found that a monocytosis- and tumour-induced anti-inflammatory effect directed against macrophages was promptly corrected by tumour excision but recurred with metastasis. These observations indicate that the monocyte abnormalities are acquired and related to cancer progression. The anti-inflammatory effect required relatively large primary or metastatic tumour burdens. Our findings support the clinical impression that monocyte abnormalities are associated with a poor prognosis.


Haematopoietic late effects of prolonged bleomycin treatment in mice.
In two studies, haematopoietic late effects of prolonged bleomycin treatment were evaluated in mice given serial injections of 21 mg bleomycin/m2 weekly for 31 and 44 weeks, respectively. Femoral bone marrow cellularity measured at 43, 45, and 49 weeks after discontinuation of the drug in the first and after 20 weeks in the second study was found to be significantly (P less than 0.05) lower in the treated mice than in the controls. CFU-S, BFU-E, and CFU-C contents were also reduced in the treated bone marrow, but with the exception of CFU-S in the second study, differences from control values were not significant. Additional long-term bone marrow cultures performed in the second study revealed no marked changes in the marrow proliferative activity and the self-renewal of stem cells to explain the reduced marrow cellularity and stem cell content. These last findings might, therefore, be due to a decrease in femoral size with less marrow content in the treated mice, since measurements of the tibial weights in both groups showed that the bones in the treated animals were significantly (P less than 0.05) lighter than those in the controls.


Fatal Legionella pneumonia: retrospective examination of lung tissue using direct and indirect fluorescent-antibody methods.
Lung tissue sections of patients who died of pneumonia within one year were screened retrospectively for Legionnaires' disease bacteria using simultaneously a direct fluorescent antibody test (DFA) and an indirect fluorescent antibody system (IFA). The IFA has shown to provide a rapid screen for the increasing number of known Legionella species and serogroups recently. In 10/168 (6%) of our cases legionellae were the pneumonia causing agents detected with both methods. Although we should have been able to detect 25 serological variants of Legionella species and serogroups with the IFA, we found Legionella pneumophila serogroup 1 only. The IFA test has proven to be a reliable means for the screening of lung tissue sections.


USP17-mediated de-ubiquitination and cancer: Clients cluster around the cell cycle.
Eukaryotic cells perform a range of complex processes, some essential for life, others specific to cell type, all of which are governed by post-translational modifications of proteins. Among the repertoire of dynamic protein modifications, ubiquitination is arguably the most arcane and profound due to its complexity. Ubiquitin conjugation consists of three main steps, the last of which involves a multitude of target-specific ubiquitin ligases that conjugate a range of ubiquitination patterns to protein substrates with diverse outcomes. In contrast, ubiquitin removal is catalysed by a relatively small number of de-ubiquitinating enzymes (DUBs), which can also display target specificity and impact decisively on cell function. Here we review the current knowledge of the intriguing ubiquitin-specific protease 17 (USP17) family of DUBs, which are expressed from a highly copy number variable gene that has been implicated in multiple cancers, although available evidence points to conflicting roles in cell proliferation and survival. We show that key USP17 substrates populate two pathways that drive cell cycle progression and that USP17 activity serves to promote one pathway but inhibit the other. We propose that this arrangement enables USP17 to stimulate or inhibit proliferation depending on the mitogenic pathway that predominates in any given cell and may partially explain evidence pointing to both oncogenic and tumour suppressor properties of USP17.


Betaxolol is equivalent to carvedilol in patients with heart failure NYHA II or III: result of a randomized multicenter trial (BETACAR Trial).
In a recent COMET trial (Lancet 362, 2003:7-13) it could be demonstrated that carvedilol < or = 50 mg/d was superior to metoprolol tartrate < or = 100 mg/d in the treatment of heart failure patients NYHA II-IV. It was investigated whether the superiority of carvedilol with its beta1, beta2 and alpha-blocking potency will persist in a comparison to a highly selective beta1-blocker with a long plasma half-time such as betaxolol. 255 pts. with NYHA II or III heart failure were double-blind randomized and uptitrated to either carvedilol 25 mg bid (n=131) or betaxolol 20 mg od (n=124). Within 8 months left ventricular ejection fraction (LVEF) increased to the same extent from 30% to 43% (carvedilol) or 31% to 43% (betaxolol) as primary endpoint (ns). 13% of the carvedilol (CAR) patients versus 15% of the betaxolol patients (BET) suffered either from cardiac death or recurrent hospitalizations (cardiac death n=6 (CAR), n=2 (BET), ns). The mean increase in the 6-min walk test was 63 m with CAR and 61 m with BET and the Minnesota living with heart failure questionnaire improved in both groups (ns). Heart rate reduction was pronounced in both groups: CAR 13.1 beats/min, BET 13.6 beats/min. The long acting highly selective beta1-blocker betaxolol in an adequate dosage is not inferior to carvedilol in terms of exercise tolerance, safety and effects on left ventricular function.


Ultrasound localization microscopy.
Ultrasound Localization Microscopy (ULM) is an emerging technique that provides impressive super-resolved images of microvasculature, i.e., images with much better resolution than the conventional diffraction-limited ultrasound techniques and is already taking its first steps from preclinical to clinical applications. In comparison to the established perfusion or flow measurement methods, namely contrast-enhanced ultrasound (CEUS) and Doppler techniques, ULM allows imaging and flow measurements even down to the capillary level. As ULM can be realized as a post-processing method, conventional ultrasound systems can be used for. ULM relies on the localization of single microbubbles (MB) of commercial, clinically approved contrast agents. In general, these very small and strong scatterers with typical radii of 1-3 µm are imaged much larger in ultrasound images than they actually are due to the point spread function of the imaging system. However, by applying appropriate methods, these MBs can be localized with sub-pixel precision. Then, by tracking MBs over successive frames of image sequences, not only the morphology of vascular trees but also functional information such as flow velocities or directions can be obtained and visualized. In addition, quantitative parameters can be derived to describe pathological and physiological changes in the microvasculature. In this review, the general concept of ULM and conditions for its applicability to microvessel imaging are explained. Based on this, various aspects of the different processing steps for a concrete implementation are discussed. The trade-off between complete reconstruction of the microvasculature and the necessary measurement time as well as the implementation in 3D are reviewed in more detail, as they are the focus of current research. Through an overview of potential or already realized preclinical and clinical applications - pathologic angiogenesis or degeneration of vessels, physiological angiogenesis, or the general understanding of organ or tissue function - the great potential of ULM is demonstrated.


Anti-human ACE2 antibody neutralizes and inhibits virus production of SARS-CoV-2 variants of concern.
The global pandemic caused by SARS-CoV-2 is a major public health problem. Virus entry occurs via binding to ACE2. Five SARS-CoV-2 variants of concern (VOCs) were reported so far, all having immune escape characteristics. Infection with the current VOC Omicron was noticed in immunized and recovered individuals; therefore, the development of new treatments against VOC infections is urgently needed. Most approved mAbs treatments against SARS-CoV-2 are directed against the spike protein of the original virus and are therefore inefficient against Omicron. Here, we report on the generation of hACE2.16, an anti-ACE2 antibody that recognizes and blocks ACE2-RBD binding without affecting ACE2 enzymatic activity. We demonstrate that hACE2.16 binding to ACE2 does not affect its surface expression and that hACE2.16 blocks infection and virus production of various VOCs including Omicron BA.1 and BA.2. hACE2.16 might, therefore, be an efficient treatment against all VOCs, the current and probably also future ones.


Gene therapeutic approach for inhibiting hepatitis C virus replication using a recombinant protein that controls interferon expression.
The hepatitis C virus (HCV) is a continuing threat to public health. The systemic administration of interferon alpha with ribavirin is the only currently approved treatment. However, this treatment is associated with a wide spectrum of systemic side effects that limits its effectiveness; thus, there is an urgent need for new treatment modalities. In this study, we describe a novel anti-HCV strategy employing a recombinant transcription factor that we have engineered in such a way that NS3/4a viral protease controls its intracellular localization, thereby restoring interferon secretion specifically in cells infected with HCV. Proof-of-concept experiments validated the strategy, showing that the recombinant transcription factor was triggered to stimulate interferon promoter by NS3/4A and remained inactive in cells without NS3/4a. Using an adenovirus-associated viral vector delivery system, we found that the recombinant transcription factor inhibited HCV replication effectively in vitro in cultured cells.


A concept of the development of the mammalian rhombencephalon based on physical rotational forces.
The premammalian forms do not show the metencephalic and myelencephalic divisions of the rhombencephalon. The seven longitudinal grey columns, in relation to the seven nerve components are direct cranial uninterrupted continuations from the spinal cord. The isthmus rhombencephali is a tubular structure. The phylogenetically ancient ependyma rich in tanycytes lines the fourth ventricle. The neural canal does not open into the subarachnoid space and the ependyma never becomes directly continuous with the pia mater. On the other hand, in mammals, the rhombencephalon is divided into the metencephalon and myelencephalon. The seven longitudinal grey columns get fragmented and their cell masses undergo migrations. The isthmus loses its tubular nature and undergoes modifications. The fourth ventricle opens into the subarachnoid space through three openings. The phylogenetically ancient ependyma is retained as a continuous structure only along the inferolateral angle of the fourth ventricle. At the openings of the lateral recesses, the ependyma becomes directly continuous with the pia, a neural crest derivative. The oval bundle, the corpus trapezoideum and the myelencephalic component of the nuclei pontis show extensive migrations. The above phenomena can be explained by physical rotation forces and not merely by migrations.


Anterior and distal tunnel orientation for anatomic reconstruction of the medial patellofemoral ligament is safer in patients with open growth plates.
In patients with open growth plates, the direction of tunneling that avoids distal femoral physis (DFP) damage in anatomic reconstructions of the medial patellofemoral ligament (MPFL) has been a topic of discussion. The objective of this study was to determine the ideal orientation for anatomic reconstructions of MPFL tunneling that minimized DFP damage while avoiding breaching the intercondylar notch. Eighty magnetic resonance images of patients aged 10 through 17 were obtained, randomly sampled from the institutional database. A de novo software was developed to obtain 3D models of the distal femur and DFP. In each model, the anatomical insertion point of the MPFL was determined as defined by Stephen. A 20-mm-depth drilling was simulated, starting from the insertion point at every possible angle within a 90° cone using 5-, 6- and 7-mm drills. Physeal damage for each pair of angles and each drill size was determined. Damage was expressed as a percentage of total physis volume. Statistical analysis was conducted using Student's t test and one-way ANOVA. Maximum physeal damage (5.35% [4.47-6.24]) was obtained with the 7-mm drill when drilling 3° cephalic and 15° posterior from insertion without differences between sexes (n.s.). Minimal physeal damage (0.22% [0.07-0.37]) was obtained using the 5-mm drill aimed 45° distal and 0° anteroposterior, not affected by sex (n.s.). Considering intra-articular drilling avoidance, the safest zone was obtained when aiming 30°-40° distal and 5°-35° anterior, regardless of sex. Ideal femoral tunnel orientation, avoiding physeal damage and breaching of the intercondylar notch, was obtained when aiming 30°-40° distal and 5°-35° anterior, regardless of sex. This area is a safe zone that allows anatomic MPFL reconstruction of patients with an open physis.


Techniques for blood loss estimation in major non-cardiac surgery: a systematic review and meta-analysis.
Estimated blood loss (EBL) is an important tool in clinical decision-making and surgical outcomes research. It guides perioperative transfusion practice and serves as a key predictor of short-term perioperative risks and long-term oncologic outcomes. Despite its widespread clinical and research use, there is no gold standard for blood loss estimation. We sought to systematically review and compare techniques for intraoperative blood loss estimation in major non-cardiac surgery with the objective of informing clinical estimation and research standards. A structured search strategy was applied to Ovid Medline, Embase, and Cochrane Library databases from inception to March 2020, to identify studies comparing methods of intraoperative blood loss in adult patients undergoing major non-cardiac surgery. We summarized agreement between groups of pairwise comparisons as visual estimation vs formula estimation, visual estimation vs other, and formula estimation vs other. For each of these comparisons, we described tendencies for higher or lower EBL values, consistency of findings, pooled mean differences, standard deviations, and confidence intervals. We included 26 studies involving 3,297 patients in this review. We found that visual estimation is the most frequently studied technique. In addition, visual techniques tended to provide lower EBL values than formula-based estimation or other techniques, though this effect was not statistically significant in pooled analyses likely due to sample size limitations. When accounting for the contextual mean blood loss, similar case-to-case variation exists for all estimation techniques. We found that significant case-by-case variation exists for all methods of blood loss evaluation and that there is significant disagreement between techniques. Given the importance placed on EBL, particularly for perioperative prognostication models, clinicians should consider the universal adoption of a practical and reproducible method for blood loss evaluation. PROSPERO (CRD42015029439); registered: 18 November 2015.PROSPERO (CRD42015029439); registered: 18 November 2015.


Lessons learned from cardiovascular outcome clinical trials with dipeptidyl peptidase 4 (DPP-4) inhibitors.
Previous trials of glucose-lowering strategies in subjects with type 2 diabetes have demonstrated a beneficial effect of intensive glycemic control on microvascular complications but failed to show a clear benefit on cardiovascular complications. The findings of meta-analyses of rosiglitazone trials suggesting that rosiglitazone might increase the risk of myocardial infarction have cast doubt on the cardiovascular safety of glucose-lowering drugs. In 2008, the US Food and Drug Administration has implemented rigorous criteria to approve new glucose-lowering drugs, requiring proof of cardiovascular safety. These regulatory requirements have led to a considerable increase in the number of cardiovascular outcome trials in type 2 diabetes to ensure that newer glucose-lowering drugs are not associated with increased cardiovascular risk. Incretin-based therapies including dipeptidyl peptidase 4 (DPP-4) inhibitors, and injectable glucagon-like peptide 1 (GLP-1) receptor agonists are novel treatment options for patients with inadequate glucose control. Although DPP-4 inhibitors have shown neutral effects on risk factors for cardiovascular diseases, it remains unclear whether treatment with these new glucose-lowering agents might be associated with a reduction in cardiovascular events. The results of the three cardiovascular outcome trials comparing DPP-4 inhibitors treatment to placebo in addition to other glucose-lowering drugs have been published. All the three DPP-4 inhibitor cardiovascular outcome trials have shown non-inferiority with regard to cardiovascular safety, compared with placebo, when added to usual care. In this review, we summarize cardiovascular outcome trials of DPP-4 inhibitors, and provide an overview of these trials and their limitations.


c-fos and c-myc expression in human endothelial cells as a function of different culture conditions.
Cultured human umbilical vein endothelial cells (HEC) could be induced to express c-fos and c-myc mRNA by either serum or ECGS (endothelial cell growth supplement). Neither agonist separately could support HEC proliferation but the combination did. Expression of c-fos and c-myc mRNA in the presence of both serum and ECGS was similar to that observed after each of the two stimuli was introduced separately. c-fos and c-myc expression in cultured HEC, even if related, is not necessarily accompanied by stimulation of cell growth.


Germline variation in apoptosis pathway genes and risk of non-Hodgkin's lymphoma.
The t(14;18)(q32;q21) translocation is the most commonly observed chromosomal translocation in non-Hodgkin's lymphoma (NHL), resulting in constitutive Bcl-2 expression and apoptosis inhibition. In addition, germline variation in both BCL2L11 (BIM) and CASP9, known regulators of apoptosis, has recently been linked to NHL risk. We conducted a comprehensive evaluation of 36 apoptosis pathway genes with risk of NHL. We genotyped 226 single-nucleotide polymorphisms (SNP) from 36 candidate genes in a clinic-based study of 441 newly diagnosed NHL cases and 475 frequency-matched controls. We used principal components analysis to assess gene-level associations, and logistic regression to assess SNP-level associations. MACH was used for imputation of SNPs in BCL2L11 and CASP9. In gene-level analyses, BCL2L11 (P = 0.0019), BCLAF1 (P = 0.0097), BAG5 (P = 0.026), and CASP9 (P = 0.0022) were associated with NHL risk after accounting for multiple testing (tail strength, 0.38; 95% confidence interval, 0.05-0.70). Two of the five BCL2L11 tagSNPs (rs6746608 and rs12613243), both genotyped BCLAF1 tagSNPs (rs797558 and rs703193), the single genotyped BAG5 tagSNP (rs7693), and three of the seven genotyped CASP9 tagSNPs (rs6685648, rs2020902, and rs2042370) were significant at P < 0.05. We successfully imputed BCL2L11 and CASP9 SNPs previously linked to NHL, and replicated all four BCL2L11 and two of three CASP9 SNPs. We replicated the association of BCL2L11 and CASP9 with NHL risk at the gene and SNP level, and identified novel associations with BCLAF1 and BAG5. Closer evaluation of germline variation of genes in the apoptosis pathway with risk of NHL and its subtypes is warranted.


Development of soil health benchmarks for managed and semi-natural landscapes.
Efforts to improve soil health require that target values of key soil properties are established. No agreed targets exist but providing population data as benchmarks is a useful step to standardise soil health comparison between landscapes. We exploited nationally representative topsoil (0-15 cm) measurements to derive soil health benchmarks for managed and semi-natural environments across Great Britain. In total, 4587 soil organic matter (SOM), 3860 pH, 2908 bulk density (BD), and 465 earthworm abundance (EA) datapoints were used. As soil properties are sensitive to site-specific characteristics, data were stratified by habitat, soil type, and mean annual precipitation, with benchmarks defined as the middle 80 % of values in each distribution - yielding 135 benchmarks. BD and pH decreased with land management intensity (agriculture > semi-natural grasslands > woodlands > heathlands > wetlands), and vice versa for SOM and EA. Normalising benchmark ranges by medians revealed soil health indicator benchmark widths increased in the order: pH < BD < SOM < EA, while width increased with decreasing land management intensity. Arable and horticulture and improved grassland exhibited narrow benchmarks for SOM, pH and BD, yet the widest EA benchmark, suggesting additional drivers impact EA patterns. Upland wetlands had the widest BD benchmarks, important when determining carbon stocks. East Anglia currently possesses the largest proportions of atypical soils, including below typical SOM (19.2 %), above typical BD (17.4 %) and pH (39.1 %), and the smallest proportions of above typical SOM (2.4 %), and below typical BD (5.8 %) and pH (2.3 %). This is found even after land use, soil type and rainfall have been considered, underscoring how urgently soil health should be addressed here. Our benchmarking framework allows landowners to compare where their measured soil health indicators fall within expected ranges and is applicable to other biomes, national and multinational contexts.


Indigenous phytotherapy for genito-urinary diseases used by the Kandha tribe of Orissa, India.
Studies on ethnomedicobotany of Kandha tribe of Orissa, India, are scanty. In view of this the original ethno-botanical information and plant specimens were collected from the Kandhamal district of Orissa by visiting the area several times. The paper reports 27 plant species belonging to 24 families used in the treatment of 17 diseases under the broad heading genito-urinary diseases by the Kandhas of Orissa. The use of these plants does not necessarily imply efficacy, but it does give a list of species that can be studied pharmacologically for its active principles and bioactive effect.


Comparison of three prognostic scoring systems in a series of 146 cases of chronic myelomonocytic leukemia (CMML): MD Anderson prognostic score (MDAPS), CMML-specific prognostic scoring system (CPSS) and Mayo prognostic model. A detailed review of prognostic factors in CMML.
Although specific prognostic models for chronic myelomonocytic leukemia (CMML) exist, few are based on large series of patients. MD Anderson prognostic score (MDAPS) has been the most useful for CMML risk assessment. Due to recent emergence of CMML-specific prognostic scoring system (CPSS) and Mayo prognostic model, we compared the three scores. One hundred forty-six CMML patients diagnosed between 1998 and 2014 were retrospectively analyzed. Univariate analysis was performed to assess prognostic impact on overall survival (OS) and leukemia-free survival (LFS) of the variables composing the scores and all items showed prognostic value on OS with the exception of the presence of circulating immature myeloid cells. Regarding LFS, only CPSS variables, bone marrow blast ≥10% and an absolute monocyte count >10×109/L had an impact. When the scores were applied, all showed an impact on OS and retained their significance in multivariate analysis. By using ROC curves and C-index, CPSS showed a slightly better predictive value for mortality and leukemia transformation. Variables composing the three indexes were compared in multivariate analysis and only CPSS parameters and platelets<100×109/L retained their significance. Based on these findings, by adding platelet count to CPSS, a new score was implemented (CPSS-P) showing the best risk prediction capability in our series. This study reinforces the validity of the tested scores.


[Spine injections in athletes].
A major proportion of athletes have intermittent acute low back pain at some stage of their sport life. Some athletes develop chronic low back pain, which may be a major source of failing in sports. Facet, epidural and intradiscal steroid injections for specific pain generators in the spine show different short- and long-term results. They can improve low back pain for specific pain generators experienced by back pain patients and might be able to help some of these athletes to avoid open surgery and shorten the time until return to sports (RTS). Facet and intradiscal steroid injections provide only short-term effects in axial low back pain and should therefore be considered with caution, whereas the different forms of epidural steroid injections for sciatica caused by radicular compression seem to be a well-proven adjuvant treatment option within a conservative therapeutic regimen. An overview of the literature related to the use of different steroids has shown that non-particulate steroids provide better results regarding safety and avoidance of major complications, especially when used at the cervical spine. However, in Germany the use of these steroids is still off-label and patients have to provide their informed consent prior to injection. In summary, careful use of spinal injections within a conservative physiotherapeutic regimen seems to improve chronic back pain and, in turn, reduce the time of RTS.


A cross-sectional analysis of the muscle strength, spinal shrinkage, and recovery during a working day of military police officers.
Military personnel has a large prevalence of back pain, especially those involved in patrolling routines, as they wear heavy protective equipment. Patrolling includes long periods of sustaining the protective equipment in a sitting or in a motor vehicle (motorcycle or car). Thus, understanding spinal loading of military police officers after patrolling by car (CAR; n = 14), motorcycle (MOT; n = 14), and administrative (ADM; n = 14) routines is relevant to establish preventive strategies. The torque of the trunk and working and anthropometric characteristics were assessed to explain spinal loading using stature variation measures. Precise stature measures were performed before and after a 6 h journey (LOSS) and 20 min after a resting posture (RECOV). The trunk extensor (PTE BM-1 ) and flexor (PTF BM-1 ) muscles' isometric peak torque were measured before the working journey. The LOSS was similar between CAR and MOT (4.8 and 5.8 mm, respectively) after 6 h of patrolling. The ADM presented the lowest LOSS (2.8 mm; P < .05). No changes in RECOV between groups were observed (P > .05). Vibration may explain the greater spinal loading involved in patrolling in comparison to the ADM. A GLM analysis revealed that BMI was the only explanatory factor for stature loss. No independent variables explained RECOV. The ability of the trunk muscles to produce force did not influence LOSS or RECOV. Military police officers involved in patrolling may require greater post-work periods and strategies designed to reduce the weight of the protective apparatus to dissipate spinal loading. The external load used in patrolling is a relevant spinal loading factor.


Phenotypic, functional, and quantitative characterization of canine peripheral blood monocyte-derived macrophages.
The yield as well as phenotypic and functional parameters of canine peripheral blood monocyte-derived macrophages were analyzed. The cells that remained adherent to Teflon after 10 days of culture had high phagocytic activity when inoculated with Leishmania chagasi. Flow cytometric analysis demonstrated that more than 80% of cultured cells were positive for the monocyte/macrophage marker CD14.


A three-dimensional fractional visco-hyperelastic model for soft materials.
Soft materials have attracted widespread attention and brought a wave of novel potential applications. Accurate mechanical characterization is crucial for improving the ration design of the desired soft elements and understanding the influence of deformation properties on themselves. However, the intrinsic complexity of mixed elasticity and viscosity in terms of their material property, posing new challenges for constitutive modeling. In this work, a fractional visco-hyperelastic constitutive model is developed by introducing the fractional viscoelasticity into finite strain: two branches are incorporated where a hyperelastic spring is used to describe the equilibrium response, and a non-linear fractional dashpot is introduced to characterize the time-dependent material response by employing the 3D fractional viscoelasticity. It provides a method from the perspective of fractional mechanics to avoid the increased number of governing equations and the associated complex calibration process caused by introducing the internal variables. The results show that the proposed model can successfully describe the rate dependent mechanical behavior of soft materials as well as predict the material behavior in different deformation modes with reasonable accuracy.


Miller-Fisher syndrome after first dose of Oxford/AstraZeneca coronavirus disease 2019 vaccine: a case report.
Miller-Fisher Syndrome (MFS) is a variant of Guillain-Barré syndrome (GBS), an acute immune-mediated neuropathy, which manifests as a rapidly evolving areflex motor paralysis. This syndrome presents as a classic triad: ophthalmoplegia, areflexia, and ataxia. MFS is usually benign and self-limited. A Caucasian patient was admitted to our hospital with the flu, loss of bilateral strength in the lower limbs and upper limbs and sudden-onset ataxia 7 days after receiving a first dose of the Oxford/AstraZeneca COVID-19 vaccine. On neurological examination, the patient had Glasgow Coma Scale score of 15, with absence of meningeal signs; negative Babinski sign; grade 2 strength in the lower limbs and grade 4 strength in the upper limbs; axial and appendicular cerebellar ataxia; and peripheral facial diparesis predominantly on the right, without conjugate gaze deviation. Cerebrospinal fluid (CSF) was collected on admission, and analysis revealed albuminocytological dissociation with CSF protein of 148.9 mg/dL; leukocytes, 1; chlorine, 122; glucose, 65 mg/mL; red cells, 2; and non-reactive venereal disease research laboratory test result. The COVID-19 IgG/IgM rapid immunological test was negative. Electroneuromyography revealed a recent moderate-grade and primarily sensory and motor demyelinating polyneuropathy with associated proximal motor block. Miller-Fisher Syndrome may be related to events other than infections prior to neuropathy, as in the case reported here. The patient presented strong correlations with findings for MFS reported in the literature, such as the clinical condition, the results of electroneuromyography, and results of the CSF analysis typical for MFS. When treatment was provided as proposed in the literature, the disease evolved with improvement. Ultimately, the diagnosis of incomplete MFS was made, including acute ataxic neuropathy (without ophthalmoplegia).


Characteristics of fluorinated nitroazoles as hypoxic cell radiosensitizers.
Types of 2-nitroimidazoles and 3-nitro-1,2,4-triazoles bearing one or two fluorine atoms on their side chains were synthesized to evaluate their physicochemical properties, radiosensitizing effects, and toxicity. The reduction potential of the compounds containing one fluorine was similar to that of misonidazole (MISO), whereas that of the difluorinated compounds was slightly higher. Both mono- and difluorinated compounds had an in vitro sensitizing activity comparable to or slightly higher than that of MISO. The fluorinated 3-nitrotriazoles were almost as efficient as the 2-nitroimidazoles with the same substituent. In vivo, some of the compounds were up to twice more efficient than MISO, whereas others were as efficient as MISO. Toxicity in terms of LD50/7 in mice was quite variable depending on the side-chain structure; the amide derivatives were less toxic than MISO, whereas the alcohol and ether derivatives were more toxic. In view of the radiosensitizing effect and toxicity in vivo, at least one compound, KU-2285 (a 2-nitroimidazole with an N1-substituent of: CH2CF2CONHCH2CH2OH) has been found to be as useful a hypoxic cell sensitizer as SR-2508.


Renal Denervation vs Pharmacotherapy for Resistant Hypertension: A Meta-Analysis.
The effect of renal denervation (RD) for resistant hypertension remains controversial because of the conflicting results of finished and ongoing studies. The authors performed a meta-analysis of case-control studies to identify whether renal sympathetic denervation or pharmacotherapy (PHAR) was more effective for resistant hypertension. A systematic Internet database search of relevant papers written in English was performed. A total of nine studies met the inclusion criteria, with a total of 1096 patients. When comparing the RD group with the PHAR group, there was a significant decrease in systolic blood pressure (SBP) (weighted mean difference, -12.81 mm Hg; 95% confidence interval [CI], -22.77 mm Hg to -2.85 mm Hg; P=.01) and diastolic blood pressure (DBP) (weighted mean difference, -5.56; 95% CI, -8.15 mm Hg to -2.97 mm Hg; P<.0001). This pooled analysis shows that for patients with resistant hypertension, RD is more effective in reducing SBP and DBP than PHAR. RD may be more effective in special subgroups of patients, which needs to be identified in future investigations.


Human CD8+ CD25 + CD127 low regulatory T cells: microRNA signature and impact on TGF-β and IL-10 expression.
Regulatory T cells (Tregs) are central for maintaining immune balance and their dysfunction drives the expansion of critical immunologic disorders. During the past decade, microRNAs (miRNAs) have emerged as potent regulators of gene expression among which immune-related genes and their immunomodulatory properties have been associated with different immune-based diseases. The miRNA signature of human peripheral blood (PB) CD8+ CD25 + CD127 low Tregs has not been described yet. We thus identified, using TaqMan low-density array (TLDA) technique followed by individual quantitative real-time polymerase chain reaction (qRT-PCR) confirmation, 14 miRNAs, among which 12 were downregulated whereas two were upregulated in CD8 + CD25 + CD127 low Tregs in comparison to CD8 + CD25 - T cells. In the next step, microRNA Data Integration Portal (mirDIP) was used to identify potential miRNA target sites in the 3'-untranslated region (3'-UTR) of key Treg cell-immunomodulatory genes with a special focus on interleukin 10 (IL-10) and transforming growth factor β (TGF-β). Having identified potential miR target sites in the 3'-UTR of IL-10 (miR-27b-3p and miR-340-5p) and TGF-β (miR-330-3p), we showed through transfection and transduction assays that the overexpression of two underexpressed miRNAs, miR-27b-3p and miR-340-5p, downregulated IL-10 expression upon targeting its 3'-UTR. Similarly, overexpression of miR-330-3p negatively regulated TGF-β expression. These results highlighted an important impact of the CD8 + Treg mirnome on the expression of genes with significant implication on immunosuppression. These observations could help in better understanding the mechanism(s) orchestrating Treg immunosuppressive function toward unraveling new targets for treating autoimmune pathologies and cancer.


Dipole tracing of monkey somatosensory evoked potentials.
Current source generators (dipoles) of somatosensory evoked potentials (SEPs) in response to electrical stimulation of the median nerve in anesthetized monkeys were localized by the dipole tracing method (DT) which can approximate surface potential distributions to an equivalent dipole. The locations and latencies of dipoles estimated by DT were confirmed by recording multiple unit activities from the parietal cortices of the same monkeys investigated by DT. Loci of generators for each SEP component were: P10 and N10, area 3b; P12, areas 1 and 2; P18, area 5. The results indicate that DT could trace the successive movement of generators of SEPs from the primary somatosensory area to area 5, which has been suggested in neuroanatomical studies to be part of the hierarchical sequence of somatosensory processes.


Assessing the health impacts of PM2.5 and ozone pollution and their comprehensive correlation in Chinese cities based on extended correlation coefficient.
The coordinated control of PM2.5 and ozone pollution is becoming more and more important in the current and next stage of Chinese environmental pollution control. Existing studies are unable to provide sufficient quantitative assessments of the correlation of PM2.5 and ozone pollution to support the coordinated control of the two air pollutants. This study develops a systematic method to comprehensively assess the correlation between PM2.5 and ozone pollution, including the evaluation of the impact of two air pollutants on human health and the extended correlation coefficient (ECC) for assessing the bivariate correlation index of PM2.5-ozone pollution in Chinese cities. According to the latest studies on epidemiology conducted in China, we take cardiovascular and cerebrovascular diseases and respiratory diseases as the ozone pollution's health burden when evaluating the health impact of ozone pollution. The results show that the health impact of PM2.5 in China decreases by 25.9 % from 2015 to 2021, while the health impact of ozone increases by 11.8 %. The ECC of 335 cities in China shows an increasing-decreasing trend but has generally increased from 2015 to 2021. The study provides important support for an in-depth understanding of the correlation and development trend of Chinese PM2.5 and ozone pollution by classifying the comprehensive PM2.5-ozone correlation performances of Chinese cities into four types. China or other countries will get better environmental benefits by implementing different coordinated management approaches for different correlative types of regions based on the assessment method in this study.


Methanol fixation. An alternative to heat fixation of smears before staining.
Methanol fixation of Gram-stained smears was compared to heat fixation. Smears were prepared in duplicate from direct clinical specimens, blood culture bottles, and bacterial colonies. Results from this study show that methanol fixation is superior in every instance to heat fixation. The morphology of bacteria and tissue cells was not distorted, little or no background debris was observed, and a better Gram reaction was noted.


Development of key performance indicators for emergency departments in Ireland using an electronic modified-Delphi consensus approach.
The objective of this study was to develop a consensus among emergency medicine (EM) specialists working in Ireland for emergency department (ED) key performance indicators (KPIs). The method employed was a three-round electronic modified-Delphi process. An online questionnaire with 54 potential KPIs was set up for round 1 of the Delphi process. The Delphi panel consisted of all registered EM specialists in Ireland. Each indicator on the questionnaire was rated using a five-point Likert-type rating scale. Agreement was defined as at least 70% of the responders rating an indicator as 'agree' or 'strongly agree' on the rating scale. Data were analysed using standard descriptive statistics. Data were also analysed as the mean of the Likert rating with 95% confidence intervals (95% CIs). Sensitivity of the ratings was examined for robustness by bootstrapping the original sample. Statistical analyses were carried out using SPSS version 16.0. The response rates in rounds 1, 2 and 3 were 86, 88 and 88%, respectively. Ninety-seven potential indicators reached agreement after the three rounds. In the context of the Donabedian structure-process-outcome framework of performance indicators, 41 (42%) of the agreed indicators were structure indicators, 52 (54%) were process indicators and four (4%) were outcome indicators. Overall, the top-three highest rated indicators were: presence of a dedicated ED clinical information system (4.7; 95% CI 4.6-4.9), ED compliance with minimum design standards (4.7; 95% CI 4.5-4.8) and time from ED arrival to first ECG in suspected cardiac chest pain (4.7; 95% CI 4.5-4.9). The top-three highest rated indicators specific to clinical care of children in EDs were: time to administration of antibiotics in children with suspected bacterial meningitis (4.6; 95% CI 4.5-4.8), separate area available within EDs (seeing both adults and children) to assess children (4.4; 95% CI 4.2-4.6) and time to administration of analgesia in children with forearm fractures (4.4; 95% CI 4.2-4.7). Employing a Delphi consensus process, it was possible to reach a consensus among EM specialists in Ireland on a suite of 97 KPIs for EDs.


The Effect of Depression and Rurality on Diabetes Control.
Having depression and living in a rural environment have separately been associated with poor diabetes outcomes, but there little is known about the interaction between the 2 risk factors. This study investigates the association of depression and rurality with glycemic control in adults, as well as their interaction. This is a repeated cross-sectional study with data collected from 2010 to 2017 (n = 1,697,173 patient-year observations), comprising a near-complete census of patients with diabetes in Minnesota. The outcome of interest was glycemic control defined as hemoglobin A1c under 8%. We used a logit model with clinic-level random effects to predict glycemic control as a function of depression, patient rurality, and their interaction, adjusted for differences in observed characteristics of the patient, clinic, and patient's neighborhood. Having depression was associated with lower probability of achieving glycemic control (P < .001). Although rurality alone had no association with glycemic control, significant interactions existed between depression and rurality. Living in a small rural town mitigated the negative association between depression and glycemic control (P < .001). Although patients with depression had poorer glycemic control, living in a small rural town reduced the negative association between depression and glycemic control.


The Identification of Factors That Influence the Quality of Bypass Anastomosis and an Evaluation of the Usefulness of an Experimental Practical Scale in This Regard.
Several factors associated with interrupted and continuous suturing techniques affect the quality of bypass anastomosis. It is difficult to determine the impact of these factors during surgery. The primary objective of this study was to evaluate factors with the potential to influence the quality of bypass anastomosis using either interrupted or continuous suturing. A secondary objective was to evaluate the usefulness of a practical scale when comparing interrupted and continuous suturing techniques to improve bypass anastomosis. Interrupted (n = 100) and continuous (n = 100) suturing techniques were used in 200 end-to-side bypasses to a depth of 3 cm and were assessed by 5 neurosurgeons. Vessel closing time (P < 0.001), stitch distribution (P < 0.001), intima-intima attachment (P < 0.001), and size of the orifice (P < 0.001) had a significant impact on the quality of the bypass regardless of the suturing technique used. The suturing technique used (interrupted or continuous) and positioning of the recipient vessel (vertical or horizontal) did not significantly influence the quality of anastomosis. Using multivariate analysis, the highest statistical significance with regard to bypass quality was attributed to the large size of the orifice and intimal attachment. There were advantages and disadvantages to both suturing techniques. The scale was a practical way to measure and improve performance.


The effects of PDT in primary malignant brain tumours could be improved by intraoperative radiotherapy.
GBM has a poor survival despite surgery and chemoradiotherapy. Cytoreduction and PDT have postulated to afford better local GBM-control. However, the interaction of PDT with newer novel therapies had not been fully investigated. This study reviewed the impact of PDT in conjunction with intraoperative radiotherapy. Case note review of prospectively collected data of GBMs treated surgically by the senior author (SE). Patients received standard therapy (ST), ST+PDT or ST+PDT+IORT. ST involved maximum safe resection, PDT involved intracavity 100 J/cm² 630 nm laser and IORT involved intracavity 10-15 Gys using the PRS400®. Patients were followed up clinically and radiologically till death. There were 73 patients included in this analysis (42 males). The average age was 59years. Thirty received PDT and 43 did not. The mean survival of PDT-patients was significantly longer than those had ST alone (62.9 weeks vs. 20.6 weeks) (p=0.001). Patients < 65 year of age survived longer than those ≥ 65 year (p=0.033). IORT on its own did not make a significant difference to survival (p=0.111). However the average survival for patients who received PDT+IORT was substantially higher than those who received PDT alone (79 weeks vs. 39.7 weeks). Using a Cox regression covariant analysis, demonstrated that PDT was the only therapy to make a statistically significant difference to survival (p=0.018). PDT in high grade glioma was statistically significant therapeutic modality and its effects were further improved by IORT.


Single dose of H3 receptor antagonist--ciproxifan--abolishes negative effects of chronic stress on cognitive processes in rats.
The role of histamine neurons in stress evoked cognitive impairments remains unclear. Previous research has indicated that the blockade of H(3)-type histamine receptors may improve attention and memory in naïve rodents. The purpose of this study was to determine if ciproxifan, (cyclopropyl-(4-(3-1H-imidazol-4-yl) propyloxy) phenyl) ketone, an H(3) receptor antagonist, could alleviate cognitive deficits observed in chronically stressed rats. Specifically, we attempted to characterize the preventive action of single dose of ciproxifan (3 mg/kg, i.p.) against an impairment caused by chronic restraint stress (2 h daily for 21 days) on recognition memory tested in an object recognition task and on the long-term memory tested in a passive avoidance test. We found that administration of ciproxifan potently prevented deleterious effects of chronic restraint stress, when administered prior to learning, or immediately after learning, or before retrieval on both the recognition (p<0.001) and the passive avoidance behavior (p<0.001). These data support the idea that modulation of H(3) receptors represents a novel and viable therapeutic strategy in the treatment of stress evoked cognitive impairments.


Patient-specific in vivo right ventricle material parameter estimation for patients with tetralogy of Fallot using MRI-based models with different zero-load diastole and systole morphologies.
Patient-specific in vivo ventricle material parameter determination is important for cardiovascular investigations. A new cardiac magnetic image (CMR)-based modeling approach with different zero-load diastole and systole geometries was adopted to estimate right ventricle material parameter values for healthy and patients with Tetralogy of Fallot (TOF) and seeking potential clinical applications. CMR data were obtained from 6 healthy volunteers and 16 TOF patients with consent obtained. CMR-based RV/LV models were constructed using two zero-load geometries (diastole and systole, 2G model). Material parameter values for begin-filling (BF), end-filling (EF), begin-ejection (BE), and end-ejection (EE) were recorded for analyses. Effective Young's moduli (YM) for fiber direction stress-strain curves were calculated for easy comparisons. The mean EE YM value of TOF patients was 78.6% higher than that of the healthy group (HG). The mean end-ejection YM value from worse-outcome TOF group (WG) post pulmonary valve replacement (PVR) surgery was 59.5% higher than that from the better-outcome TOF group (BG). Using begin-filling YM and end-ejection YM as predictors and the classic logistic regression model to different better-outcome group patients from worse-outcome group patients, the areas under Receiver Operating Characteristic (ROC) curves were found to be 0.797 and 0.883 for begin-filling YM and end-ejection YM, respectively. The sensitivity and specificity 0.761 and 0.755 using end-ejection YM as the predictor. This preliminary study suggests that ventricle material stiffness could be a potential parameter to be used to differentiate BG patients from WG patients with further effort and large-scale patient data validations.


Flow dichroism, flow polarized fluorescence and viscosity of the DNA acridine complexes.
The strong binding of various acridine dyes to DNA has been studied by the measurements of flow dichroism, flow polarized fluorescence and viscosity. Negative flow dichroism and percentage change in polarized fluorescence intensity show that intercalated dye molecules are oriented rather perpendicularly to the main axis of the DNA helix, like base pairs. On the other hand, viscosity measurements show that the increase of the contour length of DNA depends on the dye structure, being much smaller in the case of dyes with bulky substituents compared to that of the other dyes. This may be attributed to the formation of the outside bound complex. Further, the introduction of bulk substituents to the acridine ring leads to a little smaller values of the reduced dichroism and intensity change of polarized fluorescence. The results may be qualitatively understood if we assume that the outside bound dye lies in the groove of the DNA helix and the plane of the dye tilts from the perpendicular direction relative to the main axis of the helix.


Organochlorine pesticide bioaccumulation in wild Nile crocodile (Crocodylus niloticus) fat tissues: Environmental influences on changing residue levels and contaminant profiles.
Biologically significant concentrations of organochlorine pesticides (OCPs) continue to be reported in wildlife populations and are of particular concern in species that occupy the highest trophic levels. Nile crocodiles (Crocodylus niloticus) are important apex predators occurring throughout much of tropical and subtropical sub-Saharan Africa, where they inhabit estuarine and freshwater habitats often impacted by contamination. In this study we examined pesticide residue accumulation in fat tissue from Nile crocodiles at Lake St Lucia, South Africa, where historically large quantities of OCPs have been used for agriculture and disease control. During 2019, we collected tail fat samples from wild (n = 21) and captive (n = 3) individuals to examine the influence of habitat, body size and sex on variations in bioaccumulation. The principal contaminant found was p,p'-DDE, a major persistent metabolite of DDT, which continues to be used in the region for combating malaria. Tissue p,p'-DDE concentrations in wild crocodiles (95-1200 ng g-1 ww) were significantly (p < 0.05) higher compared to captive individuals (23-68 ng g-1 ww) and strongly correlated (R2 > 0.70) to body length. Male (n = 14) and female (n = 7) wild crocodiles exhibited similar contaminant body burdens, however, total concentrations were substantially lower than those measured in the same population during 2016/2017. Marked differences in residue levels and profiles appear to reflect changes in food availability and dietary exposure associated with a shift in environmental conditions. These findings suggest that periods of environmental stress may be associated with enhanced toxicological risk in crocodiles. Additional work is needed to better understand contaminant accumulation and elimination mechanisms in crocodiles, and their potential effects on reproductive health.


Disparities in uptake of cholecystectomy for idiopathic pancreatitis: A nationwide retrospective cohort study.
The majority of cases of idiopathic acute pancreatitis (IAP) are thought to result from occult biliary disease. A growing body of evidence suggests that cholecystectomy for IAP reduces the risk of recurrence by up to two thirds. This study examined nationwide uptake and disparities in adoption of cholecystectomy for IAP. The National Inpatient Sample was queried to identify admissions for IAP between October 2015 and December 2018. Patients who underwent cholecystectomy before discharge and those that did not were compared using Wald χ2 tests for categorical variables and Student's t test for continuous variables. Patient- and hospital-level predictors of cholecystectomy were identified using weighted multivariable logistic regression. Of 62,305 estimated admissions for IAP, only 665 (1.1%) underwent cholecystectomy before discharge. Female sex, initiation of total parenteral nutrition (TPN), insurance status, and hospital type were associated with cholecystectomy on univariable analysis. On multivariable analysis, Hispanic patients (odds ration [OR] 1.60, 95% confidence interval [CI] 1.01-2.56), patients on TPN (OR 2.70, 95% CI 1.17-6.24), and those with private insurance (OR 2.18, 95% CI 1.48-3.21 versus Medicare/Medicaid) were more likely to receive operations. Small hospitals and hospitals in rural areas were least likely to perform empiric cholecystectomies. Despite increasing evidence supporting cholecystectomy after IAP, the practice remains rare in the United States. Educational efforts and active implementation efforts are needed to promote adoption. Particular attention should be focused on small, rural centers and those that disproportionately care for uninsured patients and patients with public insurance.


Primary malignant tumors of the stomach in children.
Primary malignant tumors of the stomach are rare in children. During the years 1952--1977, eight children were seen at Childrens Hospital of Los Angeles with primary malignant tumors of the stomach. The patients were between the ages of 20 mo and 17 hr and included 5 males and 3 females. The series consisted of 3 malignant lymphomas, 2 adenocarcinomas, 2 leiomyosarcomas, and 1 rhabdomyosarcoma of the stomach. Three patients are living and five are dead. Clinical histories, treatment and pathologic findings are described along with a review of the literature and recommendations for the management of primary gastric malignancies in children.


Analysis of wheeze sounds during tidal breathing according to severity levels in asthma patients.
Objective: This study aimed to statistically analyze the behavior of time-frequency features in digital recordings of wheeze sounds obtained from patients with various levels of asthma severity (mild, moderate, and severe), and this analysis was based on the auscultation location and/or breath phase. Method: Segmented and validated wheeze sounds were collected from the trachea and lower lung base (LLB) of 55 asthmatic patients during tidal breathing maneuvers and grouped into nine different datasets. The quartile frequencies F25, F50, F75, F90 and F99, mean frequency (MF) and average power (AP) were computed as features, and a univariate statistical analysis was then performed to analyze the behavior of the time-frequency features. Results: All features generally showed statistical significance in most of the datasets for all severity levels [χ2 = 6.021-71.65, p < 0.05, η2 = 0.01-0.52]. Of the seven investigated features, only AP showed statistical significance in all the datasets. F25, F75, F90 and F99 exhibited statistical significance in at least six datasets [χ2 = 4.852-65.63, p < 0.05, η2 = 0.01-0.52], and F25, F50 and MF showed statistical significance with a large η2 in all trachea-related datasets [χ2 = 13.54-55.32, p < 0.05, η2 = 0.13-0.33]. Conclusion: The results obtained for the time-frequency features revealed that (1) the asthma severity levels of patients can be identified through a set of selected features with tidal breathing, (2) tracheal wheeze sounds are more sensitive and specific predictors of severity levels and (3) inspiratory and expiratory wheeze sounds are almost equally informative.


Special review series on 3D organotypic culture models: Introduction and historical perspective.
Three dimensional (3D) organ-like (organotypic) culture models are a rapidly advancing area of in vitro biological science. In contrast to monolayer cell culture methods which were developed to achieve proliferation of animal cells in the beginning of in vitro biology, the advancements in 3D culture methods are designed to promote cellular differentiation, and to achieve in vivo-like 3D structure and organotypic functions. This project was conceived through the Society for In Vitro Biology to draw on the expertise of individual scientists with special expertise in organotypic cultures of selected tissues or associated interrogation methods to prepare individual-focused reviews in this series. This introductory manuscript will review the early achievements of animal cell culture in monolayer culture and the limitations of that approach to reproduce functioning organ systems. Among these are the nature and 3D architecture of the substrate on which or in which the cells are grown, physical and mechanical clues from the substrate, cell-cell interactions, and defined biochemical factors that trigger the induction of the 3D organotypic differentiation. The organoid culture requires a source of cells with proliferative capacity (ranging from tissue-derived stem or immortalized cells to the iPSC cultures), a suitable substrate or matrix with the mechanical and stimulatory properties appropriate for the organotypic construct and the necessary stimulation of the culture to drive differentiation of the cell population to form the functioning organotypic construct. Details for each type of organotypic construct will be provided in the following papers.


Novel iridoids from the flowers of Campsis grandiflora.
A non-glycosidic iridoid, campsinol (1), and two iridoid glucosides, 7-O-(Z)-p-coumaroylcachineside V (2) and 7-O-(E)-p-coumaroylcachineside I (3), were isolated from the fresh flowers of Campsis grandiflora along with five known iridoid glycosides, ixoroside (4), campsiside (5), cachineside I (6), 5-hydroxycampenoside (7), and 5-hydroxycampsiside (8), and two known phenylpropanoid glycosides, acteoside (9) and leucosceptoside A (10). The structures of these compounds were determined based on the NMR and Mass spectroscopic data and other chemical evidences.


Toll-like receptor 3 and STAT-1 contribute to double-stranded RNA+ interferon-gamma-induced apoptosis in primary pancreatic beta-cells.
Viral infections and local production of cytokines probably contribute to the pathogenesis of Type 1 diabetes. The viral replicative intermediate double-stranded RNA (dsRNA, tested in the form of polyinosinic-polycytidylic acid, PIC), in combination with the cytokine interferon-gamma (IFN-gamma), triggers beta-cell apoptosis. We have previously observed by microarray analysis that PIC induces expression of several mRNAs encoding for genes downstream of Toll-like receptor 3 (TLR3) signaling pathway. In this report, we show that exposure of beta-cells to dsRNA in combination with IFN-alpha, -beta, or -gamma significantly increases apoptosis. Moreover, dsRNA induces TLR3 mRNA expression and activates NF-kappaB and the IFN-beta promoter in a TRIF-dependent manner. dsRNA also induces an early (1 h) and sustained increase in IFN-beta mRNA expression, and blocking IFN-beta with a specific antibody partially prevents PIC plus IFN-gamma-induced beta-cell death. On the other hand, dsRNA plus IFN-gamma does not induce apoptosis in INS-1E cells, and expression of TLR3 and type I IFNs mRNAs is not detected in these cells. Of note, disruption of the STAT-1 signaling pathway protects beta-cells against dsRNA plus IFN-gamma-induced beta-cell apoptosis. This study suggests that dsRNA plus IFN-gamma triggers beta-cell apoptosis by two complementary pathways, namely TLR3-TRIF-NF-kappaB and STAT-1.


Rapidly progressive primary leptomeningeal atypical teratoid/rhabdoid tumor: a report of 2 cases.
Atypical teratoid/rhabdoid tumor is a rare, highly malignant central nervous system tumor most commonly occurring in very young children. Atypical teratoid/rhabdoid tumor most often presents as an expanding mass with symptoms consistent with the location of the tumor and may present with metastatic leptomeningeal disease. The authors describe 2 cases of rapidly progressive, diffuse leptomeningeal atypical teratoid/rhabdoid tumor without a solid primary mass. These cases demonstrate a clinical picture that can easily be confused with a basilar meningitis, encephalomyelitis, or vasculitis.


Immunomodulatory mechanism of a purified polysaccharide isolated from Isaria cicadae Miquel on RAW264.7 cells via activating TLR4-MAPK-NF-κB signaling pathway.
The immunomodulatory effect of a novel purified polysaccharide (JCH-1) isolated from Isaria cicadae Miquel had been confirmed to promote secretion of nitric oxide (NO), tumor necrosis factor-alpha (TNF-α) and interleukin 6 (IL-6) in our previous study. However, the immunomodulatory mechanism was still unclear. The purpose of this study was to investigate the immunomodulatory mechanism of JCH-1. Experimental data showed that JCH-1 could increase protein expression of toll-like receptor 4 (TLR4), promote the phosphorylation of mitogen-activated protein kinase (MAPK), as well as nuclear factor-kappa B (NF-κB) p65. Importantly, TLR4 inhibitor inhibited JCH-1-induced activation of MAPK-NF-κB signaling pathway, thus suppressed JCH-1-induced secretion of NO, TNF-α and IL-6. Collectively, these results indicated that JCH-1 actives RAW264.7 cells through TLR4-MAPK-NF-κB signaling pathway.


Population genetics of gene function.
This paper shows that differentiating the lifetimes of two phenotypes independently from their fertility can lead to a qualitative change in the equilibrium of a population: since survival and reproduction are distinct functional aspects of an organism, this observation contributes to extend the population-genetical characterisation of biological function. To support this statement a mathematical relation is derived to link the lifetime ratio T₁/T₂, which parameterizes the different survival ability of two phenotypes, with population variables that quantify the amount of neutral variation underlying a population's phenotypic distribution.