Antiperistaltic Transverse Coloplasty: A Salvage Procedure in Extensive Bowel and Colorectal Resections to Avoid Intestinal Failure.
After extensive small and colon resections, quality of life can be affected. We propose the antiperistaltic transverse coloplasty as a solution that allows for preservation of the transverse colon after both right and left colectomies while achieving a tension-free colorectal anastomosis slowing the transit and increasing the absorption time, resulting in better stool consistency and quality of life compared with an ileorectal anastomosis. This technique was performed in a 41-year-old woman with Goblet cell adenocarcinoma of the appendix with peritoneal metastasis. The transverse colon is rotated anticlockwise over the axis of the middle colic vessels toward the left parietocolic flank and relocated to the usual position of the descending colon. After 1 year of follow-up, the patient led a normal life without parenteral nutrition with five bowel movements per day and a weight gain of 15%. The use of an antiperistaltic transverse coloplasty may be worthwhile to perform in cases of extensive bowel resections during cytoreductive surgery leading to short-bowel syndrome to avoid a permanent stoma or intestinal failure and improve patient outcomes.


Nanoarchitectonics-based model membrane platforms for probing membrane-disruptive interactions of odd-chain antimicrobial lipids.
The use of nanoscience tools to investigate how antimicrobial lipids disrupt phospholipid membranes has greatly advanced molecular-level biophysical understanding and opened the door to new application possibilities. Until now, relevant studies have focused on even-chain antimicrobial lipids while there remains an outstanding need to investigate the membrane-disruptive properties of odd-chain antimicrobial lipids that are known to be highly biologically active. Herein, using the quartz crystal microbalance-dissipation (QCM-D) and electrochemical impedance spectroscopy (EIS) techniques, we investigated how an 11-carbon, saturated fatty acid and its corresponding monoglyceride-termed undecanoic acid and monoundecanoin, respectively-disrupt membrane-mimicking phospholipid bilayers with different nanoarchitectures. QCM-D tracking revealed that undecanoic acid and monoundecanoin caused membrane tubulation and budding from supported lipid bilayers, respectively, and were only active above their experimentally determined critical micelle concentration (CMC) values. Monoundecanoin was more potent due to a lower CMC and electrochemical impedance spectroscopy (EIS) characterization demonstrated that monoundecanoin caused irreversible membrane disruption of a tethered lipid bilayer platform at sufficiently high compound concentrations, whereas undecanoic acid only induced transient membrane disruption. This integrated biophysical approach also led us to identify that the tested 11-carbon antimicrobial lipids cause more extensive membrane disruption than their respective 12-carbon analogues at 2 × CMC, which suggests that they could be promising molecular components within next-generation antimicrobial nanomedicine strategies.


Reversible homolytic activation of water via metal-ligand cooperativity in a T-shaped Ni(ii) complex.
A T-shaped Ni(ii) complex [Tol,PhDHPy]Ni has been prepared and characterized. EPR spectra and DFT calculations of this complex suggest that the electronic structure is best described as a high-spin Ni(ii) center antiferromagnetically coupled with a ligand-based radical. This complex reacts with water at room temperature to generate the dimeric complex [Tol,PhDHPy]Ni(μ-OH)Ni[Tol,PhDHPyH] which has been thoroughly characterized by SXRD, NMR, IR and deuterium-labeling experiments. Addition of simple ligands such as phosphines or pyridine displaces water and demonstrates the reversibility of water activation in this system. The water activation step has been examined by kinetic studies and DFT calculations which suggest an unusual homolytic reaction via a bimetallic mechanism. The ΔH ‡, ΔS ‡ and KIE (k H/k D) of the reaction are 5.5 kcal mol-1, -23.8 cal mol-1 K-1, and 2.4(1), respectively. In addition to the reversibility of water addition, this system is capable of activating water towards net O-atom transfer to substrates such as aromatic C-H bonds and phosphines. This reactivity is facilitated by the ability of the dihydrazonopyrrole ligand to accept H-atoms and illustrates the utility of metal ligand cooperation in activating O-H bonds with high bond dissociation energies.


A novel Dock8 gene mutation confers diabetogenic susceptibility in the LEW.1AR1/Ztm-iddm rat, an animal model of human type 1 diabetes.
The LEW.1AR1-iddm rat, an animal model of human type 1 diabetes, arose through a spontaneous mutation within the inbred strain LEW.1AR1. A susceptibility locus (Iddm8) on rat chromosome 1 (RNO1) has been identified previously, which is accompanied by autoimmune diabetes and the additional phenotype of a variable CD3(+) T cell frequency. In the present study we characterised the Iddm8 region on RNO1 in backcross strains using the genetically divergent Brown Norway (BN) and Paris (PAR) rats. Candidate genes of the Iddm8 region were sequenced for mutation analysis. The Iddm8 region could be subdivided by single nucleotide polymorphism (SNP) analyses. In the first region, a mutation in exon 44 of the Dock8 gene was identified resulting in an amino acid exchange in the protein from glutamine to glutamate. This exchange is unique for the LEW.1AR1-iddm rat. In the second region, a SNP was detected in exon 11 of the Vwa2 gene with an exchange from arginine to tryptophan. This SNP is also present in other rat strains. The Dock8 mutation gave rise to a new type 1 diabetes rat model with very close similarity to type 1 diabetes in humans, providing a deepened insight into the impact of genes involved in diabetes development.


[Content of lipids in plasma and some cytochemical patterns of leukocytes in experimental hypercholesterolemia].
Long-term per oral administration of cholesterol into rabbits showed that the content of lipid containing leukocytes might correlate within certain limits with the concentration of blood serum lipids. Very high hyperlipidemia could inhibit the phagocytic activity of RES and, thereby, deteriorate apparently the lipophagocytosis and the lipid transport by leukocytes. At the same time, the long-term feeding of rabbits with cholesterol was shown to cause lymphocytopenia and a decrease in activity of succinate- and alpha-glycerophosphate dehydrogenases in the circulating lymphocytes.


Total pancreatectomy and islet cell autotransplantation: Definitive treatment for chronic pancreatitis.
In appropriately selected patients, total pancreatectomy and islet cell autotransplant controls pain and improves quality of life while often minimizing the development of overt diabetes. Multidisciplinary management and lifelong follow-up help to maximize the benefit of this procedure. This review highlights its history, indications, metabolic outcomes, and future directions.


Hospital cost-categories of pancreaticoduodenectomy.
In the era of cost-conscious healthcare, hospitals are focusing on costs. Analysis of hospital costs per cost-category may provide indications for potential cost saving measures in pancreaticoduodenectomy (PD). Between January 2004 and June 2005, 109 consecutive patients underwent curative PD for a pancreatic or peri-ampullary tumour. Costs were defined in different cost categories for each activity centre and were linked to the individual patient via the 'bill of activities'. Postoperative complication rate was 46.8%, postoperative pancreatic fistula (POPF) 12.8%, and mortality rate 1.8%. The overall median LOS was 17 (range 7-52) days. The length of hospital stay (LOS) was significantly (p < 0.0001) different between patients with POPF, those with other complications, and patients without complications i.e. 26 (10-36) vs. 21 (8-52) vs. 14 (7-33) days, respectively. Median hospital cost per patient was 10406 (5570-30999) euros. The total hospital costs were significantly related to the LOS (p < 0.0001). The increase of total hospital costs was influenced by the hospitalization (p < 0.0001) and medical staff (p < 0.0001) costs, but not by the cost for the operation room (p = 0.233). Postoperative complications, in particular POPF, are associated with increased LOS and higher hospital costs. Any measure to reduce the incidence and severity of complications after PD will save hospital costs.


Study of high pressure carbon dioxide on the physicochemical, interfacial and rheological properties of liquid whole egg.
In this research, the impact of High Pressure Carbon Dioxide (HPCD) on the physicochemical, interfacial and rheological properties of liquid whole egg (LWE) was investigated. HPCD treatment increased the solubility, free sulfhydryl groups and surface hydrophobicity of LWE. The highest foaming ability (186%) and foaming stability (72%) were obtained at 60 min of HPCD treatment, which were 1.3-fold and 1.4-fold those of the control group (147% and 51%, respectively). Compared with the control group, the LWE emulsion had a minimum particle size (16.3 μm) when the treatment time was 75 min, and the highest absolute value of zeta potential reached 7.66 mV when the processing time was 60 min. HPCD treatment decreased the apparent viscosity of LWE at a low shear rate. Dynamic viscoelastic characteristics indicated that both the G' and G'' moduli of LWE under the HPCD process were lower than those of the control group at any frequency.


Assessment of exposure to organophosphate insecticides during spraying in Haiti: monitoring of urinary metabolites and blood cholinesterase levels.
Measurement of blood cholinesterase activity and of the urinary metabolites of fenitrothion (p-nitrocresol) and malathion (monocarboxylic acid) was used to assess the exposure to these insecticides of workers in the Haitian malaria control programme and of residents in the sprayed houses. Cholinesterase activity was significantly reduced at the end of the working week in 3 out of 28 fenitrothion workers. Urinary levels of p-nitrocresol (PNC) in the spraymen ranged from 2.2 to 25.2 mg/l. In fenitrothion workers who had no direct contact with spraying (weighers and supervisors), the cholinesterase activity remained >/= 75% of the normal control value, and the urinary PNC levels were relatively low. Urinary malathion monocarboxylic acid (MCA) levels at the end of the working week ranged between 1.1 and 5.3 mg/l in workers using malathion and their blood cholinesterase activity remained essentially normal. In both groups of workers the cholinesterase levels improved and the urinary excretion of metabolites decreased after 2 days of rest from the spraying operations. In the residents of the sprayed houses, low concentrations of PNC and MCA were detected in the urine 1 day after spraying and measurable but reduced levels were still present after 7 days. In all these cases the cholinesterase activity remained >/= 75% of the normal control value.


Preparation of isoliquiritigenin-loaded nanostructured lipid carrier and the in vivo evaluation in tumor-bearing mice.
Isoliquiritigenin-loaded nanostructured lipid carrier (ISL-NLC) was constructed and characterized. In vivo antitumor efficacy and immuno-modulation effects of ISL-NLC were evaluated in sarcoma 180 (S180)-bearing and murine hepatoma 22 (H22)-bearing mice model through intraperitoneal (i.p.) administration. The ISL-NLC biodistribution was also investigated in H22-bearing mice. Results demonstrated that the ISL-NLC had a spherical shape with a mean size of (160.73 ± 6.08) nm and encapsulation efficiency of (96.74 ± 1.81)%. ISL released from the nanoparticles was in a sustained manner with an initial burst release. ISL-NLC significantly inhibit tumor growth at 10, 20 and 40 mg/kg levels, and inhibition rates were 75.70%, 82.27% and 83.90% in the S180-bearing mice and 71.49%, 81.11% and 85.62% in the H22-bearing mice, respectively. The biodistribution study showed that ISL concentration of ISL-NLC in tumor is higher 2.5-fold than ISL suspension. The elimination half-life (t1/2), area under the curve (AUC) and the mean residence times (MRTs) of the ISL-NLC was much longer than that of the ISL suspension. As a whole, anticancer effect of ISL encapsulated in NLC was superior to ISL in suspension on H22-bearing and S180-bearing mice at the same dose and was a dose-dependent way, and ISL-NLC improved immunity of ISL. It can be inferred that nanostructured lipid carriers are a promising carrier for cancer therapy using ISL.


Transcriptional regulation of the alpha-1 type II collagen gene by nuclear factor B/p65 and Sox9 in the chondrocytic phenotype of uterine carcinosarcomas.
Uterine carcinosarcomas (U-CSs) are considered monoclonal in origin, but little is known about the mechanisms for establishment of heterologous sarcomatous components. Here, we examine the functional roles of nuclear factor κB (NF-κB)/p65 and Sox9 in the transcriptional regulation of alpha-1 type II collagen (COL2A1), a hallmark of chondrogenesis, during morphologic change in the direction of the chondrocytic phenotype. In 32 cases of U-CS, both phosphorylated p65 and Sox9 expression were colocalized in Col2A1-positive sarcomatous components, particularly in cartilaginous elements, with strongly positive correlation (ρ = 0.72, P = .005). A positive association of Col2A1 expression between protein (immunohistochemistry) and messenger RNA (in situ hybridization) assays was evident in sarcomatous components, whereas 9 cases also showed distinct positive signals for the messenger RNA without protein expression in carcinomatous elements, probably through a posttranscriptional and/or posttranslational modulation mechanism. In the Ishikawa endometrial cancer line, overexpression of p65 could activate transcription of COL2A1 promoter-intron reporters through binding to specific NF-κB sites in the first intron, along with up-regulation of Sox9. Exogenous induction of Sox9 also caused an increase in transcription of COL2A1, in contrast to a repression of the p65-mediated COL2A1 transcription, suggesting the existence of a negative feedback loop. These data, therefore, suggest that NF-κB/p65 signaling, as well as Sox9, may contribute to changes in the morphology of U-CS cells toward the chondrocytic phenotype through modulation of COL2A1 transcription.


A model to detect interactions between roxithromycin and oral contraceptives.
Twenty-two healthy women participated in a study to determine whether roxithromycin (a new macrolide antibiotic agent) obtunds the activity of a triphasic oral contraceptive. The duration of the study was four menstrual cycles. Medication was given as follows: (1) cycle 1, no medication to demonstrate ovulation; (2) cycle 2, triphasic oral contraceptive daily to suppress ovulation; (3) cycle 3, triphasic oral contraceptive daily plus roxithromycin, 150 mg b.i.d.; and (4) cycle 4, triphasic oral contraceptive daily plus rifampin, 300 mg daily. Sonography of the ovaries was performed on day 13, and serum progesterone was measured on day 21 of each cycle. Elevated progesterone indicated ovulation. The presence of a maturing follicle supported this finding. All volunteers ovulated in the first cycle and no volunteers ovulated in the second and third cycles. However, 11 women ovulated when rifampin and the triphasic oral contraceptive were given concomitantly. The findings suggest there is no reason to believe that roxithromycin interferes with the efficacy of oral contraceptives.


Reliability, validity and the structure of the General Health Questionnaire in a Chinese context.
The 30-item General Health Questionnaire (GHQ), the 24-item Self-Reporting Questionnaire (SRQ), and a Chinese version of the Minnesota Multiphasic Personality Inventory (MMPI) were administered to 225 English-speaking Chinese. The GHQ was found to have high internal consistency as a scale and to assess 5 dimensions of psychopathology: anxiety, inadequate coping, depression, insomnia, and social dysfunctioning. As a brief screening instrument, it correlated 0.49 with the SRQ. In the 'case-non-case' classification, it yielded a concordance rate of above 75% with the SRQ. Using the MMPI modal profiles derived from classification research as criterion measures, sensitivity, specificity and correct classification rates were above 70%. Implications for future research with the GHQ are discussed.


Investigating the influence of excipient batch variation on the structure, consistency and physical stability of polysorbate 60-based topical vehicles.
Fatty alcohol-polysorbate 60-water ternary systems were used as models to represent the continuous phases of the respective semisolid oil-in-water emulsions for topical delivery of cosmetic and medicinal agents. The influence of batch variation of polysorbate 60 and fatty alcohol on structure and consistency of these systems was investigated using microscopy, rheology, differential scanning calorimetry and X-ray scattering techniques. The polysorbate 60 : cetostearyl alcohol mixed emulsifying wax showed swelling in water, that is, the lamellar repeat distance continually augmented from 93 to 125 Å with water percentage 20-90%. Cetostearyl alcohol ternary systems were thicker than cetyl alcohol ones independently of polysorbate 60 batches used. All the ternary systems showed an initial increase in consistency over the first 2 weeks of storage, which was followed by slight changes in consistency (cetostearyl alcohol systems) due to the re-allocation of polysorbate 60 molecules in the gel network or significant breakdown of structure (cetyl alcohol systems) due to the transformation of swollen α-lamellar gel phase into β, γ crystals on 25°C storage. With all fatty alcohols, the consistency of polysorbate 60 ternary system was directly dependent upon interlamellar water thickness as governed by the length and distribution of polyoxyethylene groups within polysorbate 60 molecules. In relation with the composition of polysorbate 60 batches used, the consistency of ternary systems was higher when prepared with the polysorbate 60 batch containing a greater amount of sorbitan polyoxyethylene monoesters. It was proposed that the swollen α-crystalline gel phase could be better formed by sorbitan polyoxyethylene monoesters rather than sorbitan polyoxyethylene diesters.


Parsing the heterogeneity of brain-symptom associations in autism spectrum disorder via random forest with homogeneous canonical correlation.
Autism spectrum disorder (ASD) is a highly heterogeneous developmental disorder, but the neuroimaging substrates of its heterogeneity remain unknown. The difficulty lies mainly on the significant individual variability in the brain-symptom association. T1-weighted magnetic resonance imaging data from the Autism Brain Imaging Database Exchange (ABIDE) (NTDC = 1146) were used to generate a normative model to map brain structure deviations of cases (NASD = 571). Voxel-based morphometry (VBM) was used to compute gray matter volume (GMV). Singular Value Decomposition (SVD) was employed to perform dimensionality reduction. A tree-based algorithm was proposed to identify the ASD subtypes according to the pattern of brain-symptom association as assessed by a homogeneous canonical correlation. We identified 4 ASD subtypes with distinct association patterns between residual volumes and a social symptom score. More severe the social symptom was associated with greater GMVs in both the frontoparietal regions for the subtype1 (r = 0.29-0.44) and the ventral visual pathway for the subtype3 (r = 0.19-0.23), but lower GMVs in both the right anterior cingulate cortex for the subtype4 (r = -0.25) and a few subcortical regions for the subtype2 (r = -0.31 to -0.20). The subtyping significantly improved the classification accuracy between cases and controls from 0.64 to 0.75 (p < 0.05, permutation test), which was also better than the accuracy of 0.68 achieved by the k-means-based subtyping (p < 0.01). Sample size limited the study due to the missing data. These findings suggest that the heterogeneity of ASD might reflect changes in different subsystems of the social brain, especially including social attention, motivation, perceiving and evaluation.


Wnt signaling during BMP-2 stimulation of mesenchymal chondrogenesis.
Members of both the Wnt and bone morphogenetic protein (BMP) families of signaling molecules have been implicated in the regulation of cartilage development. A key component of the Wnt signaling pathway is the cytosolic protein, beta-catenin. We have recently shown that the chondrogenic activity of BMP-2 in vitro involves the action of the cell-cell adhesion protein, N-cadherin, which functionally complexes with beta-catenin. The aim of this study is to test the hypothesis that Wnts may be involved in BMP-2 induced chondrogenesis, using an in vitro model of high-density micromass cultures of the murine multipotent mesenchymal cell line, C3H10T1/2. Expression of a number of Wnt members was detected in these cultures, including Wnt-3A and Wnt-7A, whose levels were up- and downregulated, respectively, by BMP-2. To assess the functional involvement of Wnt signaling in BMP-2 induced chondrogenesis, cultures were treated with lithium chloride, a Wnt-7A mimetic that acts by inhibiting the serine/threonine phosphorylation activity of glycogen synthase kinase-3beta (GSK-3beta). Lithium treatment significantly inhibited BMP-2 stimulation of chondrogenesis as well as GSK-3beta enzymatic activity, and decreased the levels of N-cadherin protein and mRNA. Furthermore, lithium decreased BMP-2 upregulation of total and nuclear levels of LEF-1 and beta-catenin as well as their interaction during later chondrogenesis; similarly, the interaction of beta-catenin with N-cadherin was also decreased. Interestingly, lithium treatment did not affect the ability of BMP-2 to decrease ubiquitination of beta-catenin, although it did reduce the interaction of beta-catenin with GSK-3beta during late chondrogenesis (days 9-13). We suggest that the chondro-inhibitory effect of lithium on BMP-2 induced chondrogenesis indicates antagonism between lithium-like Wnts and BMP-2 during mesenchymal condensation.


[Determination of flavonoids constituents in epimedium total flavone capsule and research on its anti-osteoporosis activity].
To develop a rapid resolution liquid chromatography (RRLC) method for the simultaneous determination of epimedoside A, epimedin A1, epimedin A, epimedin B, epimedin C, icariin, baohuosideⅡ, icarisideⅠ, sagittatoside B, 2"-O-rhamnosyl icarisideⅡ, and baohuosideⅠin epimedium total flavone capsule. At the same time, the effects of the above 11 compounds on osteogenic differentiation of MC3T3-E1 cells were investigated by detecting the content of alkaline phosphatase (AKP). The results showed that baohuoside Ⅱ had the highest activities, and both the activities of baohuoside Ⅱ and icariside Ⅰ were stronger than those of icariin.In this study, the content determination method of flavonoid glycosides was established, and the anti-osteoporosis effect of monomers was compared, providing technical support for the study of the pharmacodynamic and mechanism of Epimedium total flavone capsule.


Accuracy of the high-throughput amplicon sequencing to identify species within the genus Aspergillus.
This study characterized the accuracy of high-throughput amplicon sequencing to identify species within the genus Aspergillus. To this end, we sequenced the internal transcribed spacer 1 (ITS1), β-tubulin (BenA), and calmodulin (CaM) gene encoding sequences as DNA markers from eight reference Aspergillus strains with known identities using 300-bp sequencing on the Illumina MiSeq platform, and compared them with the BLASTn outputs. The identifications with the sequences longer than 250 bp were accurate at the section rank, with some ambiguities observed at the species rank due to mostly cross detection of sibling species. Additionally, in silico analysis was performed to predict the identification accuracy for all species in the genus Aspergillus, where 107, 210, and 187 species were predicted to be identifiable down to the species rank based on ITS1, BenA, and CaM, respectively. Finally, air filter samples were analysed to quantify the relative abundances of Aspergillus species in outdoor air. The results were reproducible across biological duplicates both at the species and section ranks, but not strongly correlated between ITS1 and BenA, suggesting the Aspergillus detection can be taxonomically biased depending on the selection of the DNA markers and/or primers.


The Non-Fibrillating N-Terminal of α-Synuclein Binds and Co-Fibrillates with Heparin.
The intrinsically disordered protein α-synuclein (aSN) is, in its fibrillated state, the main component of Lewy bodies-hallmarks of Parkinson's disease. Additional Lewy body components include glycosaminoglycans, including heparan sulfate proteoglycans. In humans, heparan sulfate has, in an age-dependent manner, shown increased levels of sulfation. Heparin, a highly sulfated glycosaminoglycan, is a relevant mimic for mature heparan sulfate and has been shown to influence aSN fibrillation. Here, we decompose the underlying properties of the interaction between heparin and aSN and the effect of heparin on fibrillation. Via the isolation of the first 61 residues of aSN, which lacked intrinsic fibrillation propensity, fibrillation could be induced by heparin, and access to the initial steps in fibrillation was possible. Here, structural changes with shifts from disorder via type I β-turns to β-sheets were revealed, correlating with an increase in the aSN1-61/heparin molar ratio. Fluorescence microscopy revealed that heparin and aSN1-61 co-exist in the final fibrils. We conclude that heparin can induce the fibrillation of aSN1-61, through binding to the N-terminal with an affinity that is higher in the truncated form of aSN. It does so by specifically modulating the structure of aSN via the formation of type I β-turn structures likely critical for triggering aSN fibrillation.


A patient with genetically confirmed myoclonus-dystonia responded to anticholinergic treatment and improved spontaneously.
The various medical treatments applied to myoclonus-dystonia patients with a mutation of the ε-sarcoglycan gene (SGCE) have not been beneficial in most cases. Most patients experience progressive deterioration or static clinical courses, with only rare cases of spontaneous remission. A 19-year-old girl presented with a 14-year history of myoclonus and dystonia that severely affected her left arm, neck, and trunk. Genetic studies showed a mutation in SGCE [deletion in exon 6 (c.771_772delAT, Cys258X)]. Both myoclonus and dystonia responded to anticholinergic treatment for 7 years and improved spontaneously. The possibility of spontaneous improvement should be kept in mind when considering the therapeutic strategy in myoclonus-dystonia patients, especially when contemplating deep-brain stimulation.


A male-derived nonribosomal peptide pheromone controls female schistosome development.
Schistosomes cause morbidity and death throughout the developing world due to the massive numbers of eggs female worms deposit into the blood of their host. Studies dating back to the 1920s show that female schistosomes rely on constant physical contact with a male worm both to become and remain sexually mature; however, the molecular details governing this process remain elusive. Here, we uncover a nonribosomal peptide synthetase that is induced in male worms upon pairing with a female and find that it is essential for the ability of male worms to stimulate female development. We demonstrate that this enzyme generates β-alanyl-tryptamine that is released by paired male worms. Furthermore, synthetic β-alanyl-tryptamine can replace male worms to stimulate female sexual development and egg laying. These data reveal that peptide-based pheromone signaling controls female schistosome sexual maturation, suggesting avenues for therapeutic intervention and uncovering a role for nonribosomal peptides as metazoan signaling molecules.


Application of Adaptive DP-optimality to Design a Pilot Study for a Clotting Time Test for Enoxaparin.
Dosing of enoxaparin, like other anticoagulants, may result in bleeding following excessive doses and clot formation if the dose is too low. We recently showed that a factor Xa based clotting time test could potentially assess the effect of enoxaparin on the clotting system. However, the test did not perform well in subsequent individuals and effectiveness of an exogenous phospholipid, Actin FS, in reducing the variability in the clotting time was assessed. The aim of this work was to conduct an adaptive pilot study to determine the range of concentrations of Xa and Actin FS to take forward into a proof-of-concept study. A nonlinear parametric function was developed to describe the response surface over the factors of interest. An adaptive method was used to estimate the parameters using a D-optimal design criterion. In order to provide a reasonable probability of observing a success of the clotting time test, a P-optimal design criterion was incorporated using a loss function to describe the hybrid DP-optimality. The use of adaptive DP-optimality method resulted in an efficient estimation of model parameters using data from only 6 healthy volunteers. The use of response surface modelling identified a range of sets of Xa and Actin FS concentrations, any of which could be used for the proof-of-concept study. This study shows that parsimonious adaptive DP-optimal designs may provide both precise parameter estimates for response surface modelling as well as clinical confidence in the potential benefits of the study.


Effects of inorganic chlorine source on dioxin formation using fly ash from a fluidized bed incinerator.
Chlorine source is indispensable for polychlorinated dibenzo-p-dioxin and furan (PCDD/F) formation during municipal solid waste (MSW) incineration. Inorganic chlorine compounds were employed in this study to investigate their effects on PCDD/F formation through heterogeneous synthesis on fly ash surfaces. A fly ash sample obtained from a fluidized bed incinerator was sieved to different size fractions which served as the PCDD/F formation sources. The capability of different metal chlorides which facilitate the formation of PCDDs/Fs was found to follow the trends: Na < Mg < K < Al < Ca, when two particle fractions of >177 miocrom and 104-125 microm were used in the experiments. However, the capability of NaCl, MgCl2 and KCl did not seem much different from each other, whereas CaCl2 and AlCl3 were much more active in PCDD/F formation. NaCl and MgCl2 were relatively effective to produce more PCDDs, while KCl, AlCl3 and CaCl2 generated more PCDFs during heterogeneous reactions occurring on fly ash. 2,3,7,8-TCDF was the most significant contributor to the toxicity of the PCDDs/Fs formed from inorganic chlorine sources. Decreasing the sizes of fly ash particles led to more active formation of PCDDs/Fs when NaCl was used as inorganic chlorine in the experiment. The highest PCDDs/Fs produced from particles with size <37 microm, while the lowest PCDDs/Fs produced from particles with size >177 microm. The toxicity generally increased with decreasing size of the fly ash particles. The formation of PCDDs was mainly facilitated by the two size fractions, 104-125 microm and <37 microm, while the formation of PCDFs was favored by the two other size fractions, >177 microm and 53-104 microm.


Complete 1H and 13C NMR assignments of aurasperone A and fonsecinone A, two bis-naphthopyrones produced by Aspergillus aculeatus.
Complete assignments of 1H and 13C NMR chemical shifts of the polyketides aurasperone A and fonsecinone A were made by means of nuclear Overhauser enhancement and heteronuclear NMR correlation experiments. These compounds were isolated for the first time from Aspergillus aculeatus, an endophytic fungus obtained from leaves of Melia azedarach(Meliaceae).


Molecular design and experimental study of cellulose conversion to 5-hydroxymethylfurfural catalyzed by different ratios of Brønsted/Lewis acid ionic liquids.
Cellulose conversion into 5-hydroxymethylfurfural (5-HMF) is difficult because of the strong hydrogen bonding existed in cellulose chains. Brønsted/Lewis (B/L) biacidic functionalized ionic liquids (ILs) have great advantages in acid-catalyzed tandem reactions, but the catalytic effect of ILs differs considerably depending on B/L acid ratios. Therefore, this work designed a series of reactions with different proportions of biacidic ILs for the preparation of 5-HMF from cellulose. The tandem reaction is often performed in the presence of a solvent, and the activity of the catalyst is also affected by the solvent. Therefore, in this work, the solvation model density(SMD) model was introduced into the quantum chemical calculation method for molecular design to predict the catalytic effect and explore the catalytic mechanism. The calculation results and experiments jointly showed that [(HSO3-P)2im]Cl·ZnCl2 had the highest efficiency, with a 5-HMF yield of 65.66%. This study facilitates the directional optimization design of the catalyst.


Molecular determinants of intrinsic cellular stiffness in health and disease.
In recent years, the role of intrinsic biophysical features, especially cellular stiffness, in diverse cellular and disease processes is being increasingly recognized. New high throughput techniques for the quantification of cellular stiffness facilitate the study of their roles in health and diseases. In this review, we summarized recent discovery about how cellular stiffness is involved in cell stemness, tumorigenesis, and blood diseases. In addition, we review the molecular mechanisms underlying the gene regulation of cellular stiffness in health and disease progression. Finally, we discussed the current understanding on how the cytoskeleton structure and the regulation of these genes contribute to cellular stiffness, highlighting where the field of cellular stiffness is headed.


Self-association of lysozyme. Thermochemical measurements: effect of chemical modification of Trp-62, Trp-108, and Glu-35.
Heats of dilution and of saccharide binding for hen egg white lysozyme have been measured at 30 degrees, 0.1 ionic strength, and pH 7 over the range 3 to 95 mg of protein/ml. The concentration dependence of the apparent relative molar enthalpy of lysozyme derived from these results gives the thermodynamic parameters for the formation of an intermolecular contact in an indefinite (head-to-tail) self-association process as delta G 0 = -3.9 kcal/mol, delta H 0 = -6.4 kcal/mol, and delta S 0 = -8,3 e.u. Oxindolealanine-62-lysozyme does not undergo self-association reactions that can be detected calorimetrically. This derivative reacts with native lysozyme to form hybrid polymeric species with free energy and enthalpy of interaction similar to those for the polymers of native lysozyme. These results are consistent with the intermolecular contact in the self-assocaition of lysozyme being asymmetric (head-to-tail). The heat of dilution of the derivative of lysozyme in which Glu-35 is blocked as the ester with oxindolealanine-108 is like that observed for native lysozyme in acid solution and is independent of pH. The concentration difference spectrum that develops through self-association is of the shape expected for introduction of an indole chromophore into a charge-free region of the intermolecular contact. The foregoing results indicate that Glu-35 and Trp-62 are part of the contact, that perturbation of Trp-108 does not make a principle contribution to the concentration difference spectrum, and that no acid group other than Glu-35 is perturbed by self-association. There is a small change in the heat of (GlcNAc)3 binding over the range 0.005 to 0.034 M saccharide. These data give the value of -1 kcal/mol for the enthalpy change for formation of the 2:1 saccharide-enzyme complex (ES2) from ES and S.


General expression profiles of human native odontoblasts and pulp-derived cultured odontoblast-like cells are similar but reveal differential neuropeptide expression levels.
Odontoblasts play a central role during the dentin formation by organic matrix production and mineralisation. Recently, suitable in vitro techniques for studying mature primary odontoblasts and the newly differentiated odontoblasts have been developed. Firstly, the gene expression profiles of native and cultured odontoblasts were compared at large-scale to investigate the similarities and differences between the samples. Secondly, differential expression levels of the genes encoding neuronal proteins were analyzed to study odontoblasts sensory function. Microarray analysis was performed to mature native and cultured pulp-derived odontoblast-like cells to compare their transcriptome. Then, the probes positive only in one sample were divided into gene ontology categories. Expression levels of selected neuronal proteins were further studied with quantitative PCR, and at the protein level by immunofluorescence of mature and newly differentiated odontoblasts in developing tooth. Remarkable similarities between the general and neuronal protein gene expression profiles were observed. Higher cortistatin, galanin, somatostatin receptor 1 (SSTR1) and tyrosine phosphatase receptor type Z1 (PTPRZ1) expression was detected in native than in cultured odontoblast at the mRNA level. Pronociceptin was more abundantly expressed in cultured than in native odontoblasts. Immunofluorescence of mature and newly differentiated odontoblasts on human tooth germs confirmed the results. Cultured odontoblasts used in this study have similar general gene expression pattern to native odontoblasts, and therefore offer a valuable tool for the in vitro odontoblast studies. The expression of PTPRZ1 and galanin, which participate in sensory signal transduction, supports the previously suggested role of odontoblasts as sensory cells.


Healthy Eating Policy: Racial Liberalism, Global Connections and Contested Science.
The challenges to designing and implementing ethically and politically meaningful eating policies are many and complex. This article provides a brief overview of Anne Barnhill and Matteo Bonotti's Healthy Eating Policy and Political Philosophy: A Public Reason Approach while also critically engaging with the place of racial justice, global interconnectedness, and debates over science in thinking about ethics and politics of public health nutrition and policy. I do not aim to burden Barnhill and Bonotti with the responsibility to fully address these issues, but considering the interconnection of these issues and the ever pressing effects of climate change on local and global food systems, we collectively need to turn to these difficult and pressing questions about what a just food system looks like, what concerns are centred, and who is left out. I group these engagements with Barnhill and Bonotti under three headings: racial liberalism, global food system, and contested nutrition science. I conclude with some remarks about locality.


Direct photo-patterning of hyaluronic acid baits onto a fouling-release perfluoropolyether surface for selective cancer cell capture and immobilization.
A simple photolithographic process for directly patterning glycidyl methacrylate modified hyaluronic acid features onto UV curable perfluoropolyether-based surfaces is presented. Due to the versatility of the developed method, HA spotted areas with different geometrical features could be rapidly and inexpensively designed. In addition, the excellent antifouling and fouling-release properties of the substrates enabled direct HA baits photo-grafting onto PFPEs without further surface passivation or chemical modification to avoid not specific adsorption. The aim of the study was to locally switch the surface properties of the PFPEs from cells and protein repulsive to adherent. Particularly, we exploited HA well-known preferential interactions with CD44 transmembrane receptors to selectively immobilize cancer cells. Living cell arrays offer a higher-resolution visualization of HA-CD44 interactions and may provide a deep insight into understanding molecular mechanisms needed to develop selective therapies and diagnosis against tumor growth.


Absence of functional RNA encoded by a silent chromosome in non-complementing diploids obtained from protoplast fusion in Bacillus subtilis.
The molecular basis for lack of phenotypic expression of one chromosome in Bacillus subtilis non-complementing diploid clones (Ncd cells) was investigated. Correlations between chromosomal inactivation and absence of functional transcripts were determined with wild-type prophage phi 105 or a thermoinducible mutant phi 105 cts23, on either the expressed or the silent chromosome. It appears that no significant amount of phage mRNA is detectable in Ncd cells carrying the prophage in the inactive chromosome. However, phi 105 mRNA represents 0.23% of total cellular mRNA in an Ncd strain with the prophage in the expressed chromosome and 0.28% in the parental lysogenic strain. The lack of an mRNA repressor of phi 105 prophage from the silent chromosome was confirmed by the absence of repressor activity in Ncd clones with a temperature sensitive mutant phi 105 located in the silent chromosome. After heat induction, no phage production was observed. As expected these clones do not exhibit phi 105 immunity when superinfected with the same phage. The combined data of the present and previous work suggest that control of phenotypic suppression of Ncd strains should, at the transcription level, involve a different DNA tertiary organisation in one of the two chromosomes.


Hypocalcemia and hypophosphatemia in acutely ill patients.
Hypocalcemia and hypophosphatemia have recently been recognized as common metabolic complications in acutely ill patients. The diagnostic and therapeutic approaches to these abnormalities are described in this article.


Temperature changes of renal calyx during high-power flexible ureteroscopic Moses holmium laser lithotripsy: a case analysis study.
The risk of thermal damage increases with the introduction of high-power lasers during holmium laser lithotripsy. This study aimed to quantitatively evaluate the temperature change of renal calyx in the human body and the 3D printed model during high-power flexible ureteroscopic holmium laser lithotripsy and map out the temperature curve. The temperature was continuously measured by a medical temperature sensor secured to a flexible ureteroscope. Between December 2021 and December 2022, willing patients with kidney stones undergoing flexible ureteroscopic holmium laser lithotripsy were enrolled. High frequency and high-power settings (24 W, 80 Hz/0.3 J and 32 W, 80 Hz/0.4 J) were performed for each patient with room temperature (25 °C) irrigation. In the 3D printed model, we studied more holmium laser settings (24 W, 80 Hz/0.3 J, 32 W, 80 Hz/0.4 J and 40 W, 80 Hz/0.4 J) with warmed (37 °C) and room temperature (25 °C) irrigation. Twenty-two patients were enrolled in our study. With 30 ml/min or 60 ml/min irrigation, the local temperature of the renal calyx did not reach 43 °C in any patient under 25 °C irrigation after 60 s laser activation. There were similar temperature changes in the 3D printed model with the human body under the irrigation of 25 °C. Under the irrigation of 37 °C, the temperature rise slowed down, but the temperature in the renal calyces was close to or even exceeded the 43 °C at the setting of 32 W, 30 ml/min and 40 W, 30 ml/min after continuing laser activation. In the irrigation of 60 ml/min, the temperature in the renal calyces can still be maintained within a safe range after continuous activation of a holmium laser up to 40 W. However, continuous activation of 32 W or higher power holmium laser in the renal calyces for more than 60 s in the limited irrigation of 30 ml/min can cause excessive local temperature, in such situation room temperature perfusion at 25 ℃ may be a relatively safer option.


Metatarsal fracture as a consequence of foot dystonia in Parkinson's disease.
We report a 45-year-old man with a 4-year history of Parkinson's disease complicated by the development of left-foot dystonia resulting in a fracture of the fifth metatarsal. Orthopedic injuries are common in Parkinson's disease, but they are usually secondary to falls. Although drug-induced dystonia is a common side effect of pharmacological treatment of Parkinson's disease, this is the first report of a fracture related to these abnormal movements.


Oil-based adjuvants delivered intradermally induce high primary IgG2 immune response in swine.
The effects of intradermal application of antigen with or without different adjuvants and activation of immune response are presented in this study. Six groups of six piglets each were immunized with keyhole limpet haemocyanin (KLH) antigen in combination with aluminium hydroxide or oil-based adjuvants (complete and incomplete Freund's adjuvants, Montanide ISA 206 and Emulsigen). IgG1 and IgG2 levels in sera were measured by KLH-specific ELISA. Interestingly, oil-based adjuvants induced high primary IgG2 response, suggesting the Th1 lymphocyte polarization. Also, considering the similarities between human and porcine organism, we suggest that intradermal application could be considered as an effective vaccine delivery route in both veterinary and human medicine.


Renormalization of metabolic coupling treats age-related degenerative disorders: an oxidative RPE niche fuels the more glycolytic photoreceptors.
Retinitis pigmentosa is characterized by a dysregulation within the metabolic coupling of the retina, particularly between the glycolytic photoreceptors and the oxidative retina pigment epithelium. This phenomenon of metabolic uncoupling is seen in both aging and retinal degenerative diseases, as well as across a variety of cell types in human biology. Given its crucial role in the health and maintenance of these cell types, the metabolic pathways involved present a suitable area for therapeutic intervention. Herein, this review covers the scope of this delicate metabolic interplay, its dysregulation, how it relates to the retina as well other cell types, and finally concludes with a summary of various strategies aimed at reinstating normal metabolic coupling within the retina, and future directions within the field.


Precision medicine is coming to town: personalising home ventilatory equipment in COPD patients with chronic hypercapnic respiratory failure.
Discharging a chronic critically ill patient is a risky procedure if the clinician does not have full control of his prescription. This is even more important when applying a machine to replace a failing organ, as is the case for home ventilation. Even if modern home ventilators fulfil quality and safety criteria and, 'on paper', ventilators and masks look very similar, performance and scenarios of applicability are not always equivalent. In the case of ventilators, the type of circuit, accessories provided and available modes vary between devices. Bench studies comparing ventilators have shown large differences in triggering, rise time, pressurisation capacities, maximal flow provided, cycling and level of authorised expiratory positive airway pressure. Automated algorithms to deal with leaks also vary and have not been sufficiently evaluated. In the case of interfaces, the choice of mask requires careful evaluation of the underlying disease and of the type of ventilator and circuit, which could have a potentially major impact on patient compliance and clinical effectiveness. This could explain different results in the same clinical situation. The choice of ventilator and type of mask represents a medical prescription and should be respected by the provider and not subject to financial constraints.


Mild leptospirosis with three-year persistence of IgG- and IgM-antibodies, initially manifesting as carpal tunnel syndrome.
Long-term persistence of IgG- and IgM-antibodies against leptospira after mild leptospirosis has not been reported. In a 45-year-old female pet-shop worker with carpal tunnel syndrome, accompanied by arthralgias, coughing, repeatedly elevated temperature, followed by easy fatigability, personality change, memory and speech disturbance, blurred vision, myalgia and swollen lymph nodes, leptospirosis was diagnosed, based upon history, clinical findings, and serological investigations. After the described symptoms had disappeared following doxycyclin for 2 weeks, IgG- and IgM-antibodies against leptospira remained positive during the next three years. This case illustrates that leptospirosis may start as carpal tunnel syndrome and that the severity of leptospirosis does not seem to be related to the intensity of the humoral immune response against the causative agent.


The linker-protein network: control of nucleosomal DNA accessibility.
Numerous studies have recently addressed the accessibility of nucleosomal DNA to protein factors. Two popular concepts - the histone code and chromatin remodeling - consider the nucleosome as a passive entity that 'waits' to be marked by histone modifications and is 'mobilized' by ATP-dependent remodelers. Here, we propose a holistic view of the nucleosome as an active, dynamic entity, the accessibility of which is controlled by binding of different linker proteins to the DNA entry/exit site. The linker proteins might directly compete for this binding site; alternatively, protein chaperones and/or chromatin remodelers might exchange one linker protein for another. Finally, according to our proposed model, the exchange factors are themselves controlled by post-translational modifications or binding of protein partners, to respond to the ever-changing intra- and extra-cellular environment.


Lead-cadmium interaction: effects on viral-induced mortality and tissue residues in mice.
Simultaneous exposure to lead and cadmium is highly probable due to common environmental sources of exposure. Mice were exposed simultaneously for 10 wek to lead and cadmium and effects were observed on accumulation of the metals in tissues and susceptibility to viral challenge. The incidence of viral-induced mortality was enhanced in all groups of mice exposed to lead and suppressed in groups of mice treated with cadmium. Mice exposed to both lead and cadmium had mortality rates intermediate to those observed after exposure to the individual metals. Lead and cadmium residues in tissues of mice exposed to both metals, as compared to residues in tissues of mice which received lead or cadmium only, indciated that interaction occurs in regard to metabolism, storage, and excretion of these elements. Chronic coexposure to lead and cadmium appears to alter certain effects produced by exposure to each metal alone.


Cloning, characterization, and expression analysis of a putative 17 beta-hydroxysteroid dehydrogenase 11 in the abalone, Haliotis diversicolor supertexta.
The 17-beta-hydroxysteroid dehydrogenases (17β-HSDs) are key enzymes for sex steroid biosynthesis. To date, relatively little is known about the presence and function of 17β-HSDs in marine gastropods. In the present study, a cDNA sequence encoding putative 17β-HSD type 11 (17β-HSD-11) was identified in marine abalone (Haliotis diversicolor supertexta). The full-length cDNA contains 1058bp, including an open reading frame (ORF) of 900bp that encodes a protein of 299 amino acids. Comparative structural analysis revealed that abalone 17β-HSD-11 shares relatively high homology with other 17b-HSD-11 hormologues, and a lesser degree of amino acid identity with other forms of 17b-HSD, especially in the functional domains, including the cofactor binding domain (TGxxxGxG) and catalytic site (YxxSK). Phylogenetic analysis showed that abalone 17β-HSD-11 belongs to the short-chain dehydrogenase/reductase (SDR) family. Functional analysis following transient transfection of the ORF into human embryonic kidney-293 (HEK-293) cells indicated that abalone 17β-HSD-11 has the ability to convert 5α-androstane-3α,17β-diol (3α-diol) to androsterone (A) and testosterone (T) to androstenedione (4A). Expression analysis in vivo demonstrated that abalone 17β-HSD-11 is differentially expressed during three stages (non-reproductive, reproductive, and post-reproductive). Taken together, these results indicate that ab-17β-HSD-11 is an SDR family member with a potential role in steroid regulation during the reproductive stage.


Pharmacological inhibition of Na+/K+-ATPase induces neurovascular degeneration and glial cell alteration in the rat retina.
Na+/K+-ATPase (NKA) plays an important role in ion homeostasis and neurotransmitter uptake. In the retina, multidirectional communications among neurons, glia, and blood vessels (that is, neuro-glio-vascular interaction) are crucial for maintaining tissue homeostasis. We investigated the role of NKA in the elements of neuro-glio-vascular unit in neonatal and adult rat retinas. Male Sprague-Dawley rats (1- and 8-week-old) were injected intravitreally with ouabain (20 nmol/eye), an inhibitor of NKA. Morphological changes in retinal neurons, glia, and blood vessels were examined. The intravitreal injection of ouabain decreased the number of cells in the ganglion cell layer, as well as the thicknesses of the inner plexiform and inner nuclear layers in neonatal and adult rats compared to age-matched controls. The ouabain-induced neuronal cell damage was partially prevented by D-(-)-2-amino-5-phosphonopentanoic acid, an antagonist of N-methyl-D-aspartic acid receptors. In the deep retinal vascular plexus of the ouabain-injected eyes, angiogenesis was delayed in neonatal rats, whereas capillary degeneration occurred in adult rats. The immunoreactivity of glutamine synthetase and vascular endothelial growth factor (VEGF) decreased in the retinas of neonatal and adult rats injected intravitreally with ouabain. The immunoreactivity of glial fibrillary acidic protein was enhanced in the retinas of ouabain-injected adult eyes. After the ouabain injection, CD45-positive leukocytes and Iba1-positive microglia increased in the inner retinal layer of neonatal rats, whereas they increased in the middle retinal layer of adult rats. These results suggest that the inhibition of NKA induces the degeneration of neuronal and vascular cells and alteration of glial cells in both neonatal and adult retinas. In addition to the direct effects of NKA inhibition, the disturbance of retinal glutamate metabolism and decreased VEGF expression may contribute to neurovascular degeneration. The activity of NKA is crucial for maintaining elements of neuro-glio-vascular unit in the retina.


Psychometric properties of screening questionnaires to detect depression in primary healthcare setting in rural Ethiopia.
Much of the research about the validity of depression screening questionnaires is on criterion validity. Evidence is scarce on the concurrent, convergent and construct validity of these measures, particularly from low-income countries. This study aimed to evaluate the psychometric properties of depression screening questionnaires in primary healthcare (PHC) in rural Ethiopia. A facility-based cross-sectional study was conducted with 587 participants recruited from patients attending three PHC facilities and two 'Holy water sites' (places where religious treatment is being provided). The psychometric properties of five mental health screening questionnaires were evaluated: the nine item Patient Health Questionnaire (PHQ-9), the two item version of PHQ-9 (PHQ-2), a version of PHQ-9 with two added items of irritability and noise intolerance (PHQ-11), the Patient Health Questionnaire-15 (PHQ-15), and the World Health Organization-Five Well-being Index (WHO-5). Clinical diagnosis of depression was ascertained by psychiatrists. We analyzed data using exploratory factor analysis, Spearman's rank order correlation coefficient (Rho), the Mann Whitney test of the equality of medians, univariate logistic regression and Cronbach's alpha. PHQ-9, PHQ-11 and WHO-5 were found to be unidimensional, with items in each scale highly loading onto one factor (factor loadings ranging from 0.64 to 0.87). The items of each instrument were internally consistent, with Cronbach's alpha ranging from 0.72 (PHQ-2) to 0.89 (PHQ-11). Scores for all screening scales were moderately or highly correlated with each other (Rho = 0.58 to 0.98) and moderately correlated with anxiety and disability scores. Median scores of all screening scales were significantly higher in those diagnosed with depression. The association of items measuring emotional and cognitive symptoms with the diagnosis of depression was stronger than the association with items measuring somatic symptoms. Irritability and noise intolerance had higher association with depression diagnosis than PHQ-9 items. Emotional and cognitive symptoms are more useful than somatic symptoms to predict the diagnosis of depression in the PHC context in Ethiopia. Future research should focus on testing the unidimensionality of PHQ-9, PHQ-11 and WHO-5 using confirmatory factor analysis; establishing the criterion validity of PHQ-11 and WHO-5; and on assessing test-retest reliability of all the measures.


Role of molecular chaperones in biogenesis of the protein kinome.
Molecular chaperones promote polypeptide folding in cells by protecting newly made and otherwise misfolded proteins against aggregation or degradation by the ubiquitin proteasome pathway. The roles of Saccharomyces cerevisiae Cdc37 and Ydj1 molecular chaperones are described in this chapter. We focus on biogenesis of protein kinases that require several different molecular chaperones for their proper folding. Specific among these is Cdc37, which binds directly to its kinase clients either during or shortly after translation and protects them against rapid proteasomal degradation. Ydj1 has a similar role, but is less specific for protein kinases in its role as a molecular chaperone. The method that we describe uses pulse chase and immunoprecipitation to analyze the fate of newly made proteins. Two kinetically distinct pathways of degradation can be discerned using this methodology that is dependent on the presence of an Hsp90 inhibitor or occurs in mutants of the molecular chaperones under study. The first is "zero-point" degradation that occurs either during or immediately after translation. The second is a slower pathway, where the half-life of kinase is approximately 20 min after translation.


Mercury toxicity risk and corticosterone levels across the breeding range of the Yellow-breasted Chat.
Mercury (Hg) is an environmental contaminant that can negatively impact human and wildlife health. For songbirds, Hg risk may be elevated near riparian habitats due to the transfer of methylmercury (MeHg) from aquatic to terrestrial food webs. We measured Hg levels in tail feathers sampled across the breeding range of the Yellow-breasted Chat (Icteria virens), a riparian songbird species of conservation concern. We assessed the risk of Hg toxicity based on published benchmarks. Simultaneously, we measured corticosterone, a hormone implicated in the stress response system, released via the hypothalamus-pituitary-adrenal axis. To better understand range-wide trends in Hg and corticosterone, we examined whether age, sex, subspecies, or range position were important predictors. Lastly, we examined whether Hg and corticosterone were correlated. Hg levels in chats were relatively low: 0.30 ± 0.02 µg/g dry weight. 148 out of 150 (98.6%) had Hg levels considered background, and 2 (1.6%) had levels considered low toxicity risk. Hg levels were similar between sexes and subspecies. Younger chats (<1 year) had higher Hg levels than older chats (>1 year). Hg levels were lowest in the northern and central portion of the eastern subspecies' range. Corticosterone concentrations in feathers averaged 3.68 ± 0.23 pg/mm. Corticosterone levels were similar between ages and sexes. Western chats had higher levels of corticosterone than eastern chats. Hg and corticosterone were not correlated, suggesting these low Hg burdens did not affect the activity of the hypothalamus-pituitary-adrenal axis. Altogether, the chat has low Hg toxicity risk across its breeding range, despite living in riparian habitats.


Circular economy as a key for industrial value chain resilience in a post-COVID world: what do future engineers think?
While the COVID-19 pandemic has led to many disruptions in industrial value chains, the adoption of circular economy (CE) principles appears to be a commendable solution for more robust, resilient, and sustainable industrial supply chains. In this study, the standpoints and visions of two consecutive classes of engineering students - following the course "Circular Economy & Industrial Systems" at the Université Paris-Saclay - are given on how they value CE strategies to mitigate the impact of COVID-19 on industrial practices. Capturing and understanding the viewpoints of the engineers of tomorrow on such a pressing issue is key to train and provide them with the suitable methods and tools to build a more circular and sustainable society. At the end of their eight-week training class, including theoretical background on industrial ecology tools, workshops, and a hands-on project, part of the final exam included a one-hour essay in which the students had to argue their position on the following questions: (i) "Circular Economy as an answer to the COVID-19 crisis?" for the class of 2020, and (ii) "Circular Economy as an answer for green recovery and value chain resiliency in the COVID-19 context?" for the class of 2021. Interestingly, the evolution of viewpoints between the beginning of the COVID-19 crisis (exam conducted in May 2020 for the first class) and one year after (exam conducted in Mars 2021 for the second class) is discussed and illustrated. Also, the answers and insights provided by engineering students on these questions are positioned within the state-of-the-art literature on the topic. Last but not least, key recommendations and challenges on how CE could alleviate COVID-related disruptions and production shortages are synthesized in a SWOT (strengths, weaknesses, threats, and opportunities) diagram.


Indigenous participation in pediatric Indigenous health research in Canada: a systematic review.
For health inequities to be successfully addressed through health research, it is necessary for researchers to strive for genuine engagement with stakeholders. Indigenous people provide critical perspectives in Indigenous health research. The objective of this review was to systematically review the existing pediatric Indigenous health research in Canada to determine the prevalence of Indigenous participation. Embase, MEDLINE, Cochrane Library were searched on April 15, 2017 and updated on July 16, 2020. A total of 798 studies focused on the health of Indigenous children ≤18 in Canada were included, of 17,752 abstracts screened in English and French. A total of 46.1% of articles indicated Indigenous participation, increasing over time. Organization/government was the most common form of Indigenous participation (62.8%) and Indigenous researcher as author was least common (10.9%). Participation by child age, geography and topic area varied. The most common category of topic researched was nutrition, lifestyle and anthropometrics. Indigeneity of researchers was determined by self-identification in the papers and may be an underestimate. Although improving over time, less than half of studies about Indigenous children in Canada included Indigenous participation in their execution. Journals and funding bodies must ensure fulsome participation of Indigenous people in research focused on Indigenous children. IMPACT: Indigenous participation in pediatric Indigenous health research is critical to producing ethical relevant and actionable results. This review describes the status of Indigenous participation in this body of work in Canada. This review highlights areas of concern and strength to improve the practices and ethics of medical researchers in this area, thereby increasing relevance of pediatric Indigenous health research to communities.


Gelatinolytic and caseinolytic proteinase activities in human prostatic secretions.
Seminal fluid contains a number of proteinase activities, many of which are secreted by the prostate gland. Our objective was to determine proteinase activities in human prostatic secretions which can degrade gelatin and/or casein. Prostatic secretions were collected by prostate massage from men with benign prostatic hyperplasia prior to surgery to relieve obstruction. Significant proteinase activities towards gelatin of about 81, 86, 94, 111, 115 and 163 Kd as well as less active forms of 23, 36, 38, 132, 137, and 148 Kd were detected using protein substrate-polyacrylamide gel zymography. In addition, Ca2+ stimulated activities of approximately 64, 66, 71 and 76 Kd; however, EDTA and EGTA inhibited all activities but the 23, 36 and 38 Kd forms (these were inhibited by benzamidine and epsilon-amino caproic acid). This suggests that the gelatinolytic activities of 64 Kd and greater were metalloproteinases and those of 23, 36, and 38 Kd were serine proteinases. Significant caseinolytic activities of 22, 25, 35, 37, 57, 90, 96, 102 and 116 Kd were found as well as several less active forms and a 12 Kd activity stimulated by Ca2+. Caseinolytic activities of 12, 14, 16, 96, 102, 116, and 126 Kd were inhibited by EDTA and EGTA indicating they are metalloproteinases. The 35, 37, 57 and 58 Kd caseinolytic activities were inhibited by benzamidine, and the 57 and 58 Kd forms by epsilon-aminocaproic acid suggesting they were serine proteinases. There was considerable variability among individuals in the molecular forms of proteinase activity expressed as well as the level of their activity. A significant decrease in the frequency of expression of the 132 Kd gelatinolytic activity was found in secretions from men with atypia or adenocarcinoma, as compared with men with benign prostatic hyperplasia alone. Our results show that human prostatic secretion contains a variety of proteinase activities. The expression of the 132 Kd gelatinolytic activity could prove useful in further evaluation of neoplastic prostatic disease.


Kaonic nuclear systems KN and KNN as decaying states.
The formation spectra of model KN and KNN systems formed by (K(-),n) reactions are investigated in order to obtain a theoretical basis for a proper interpretation of experimental data concerning kaonic nuclear quasi-bound states. It has been clarified that the experimentally observable kaonic nuclear state K(-)pp should be regarded as the decaying state introduced by Kapur-Peierls, which is different from the pole state solution of the Faddeev equation.


A Rational Approach to Organo-Photocatalysis: Novel Designs and Structure-Property Relationships.
Organic photocatalysts are emerging as viable and more sustainable tools than metal complexes. Recently, the field of organo-photocatalysis has experienced an explosion in terms of applications, redesign of well-established systems, and identification of novel scaffolds. A rational approach to the structural modification of the different photocatalysts is key to accessing unprecedented reactivity, while improving their catalytic performances. We herein discuss the concepts underpinning the scaffold modification of some of the most recently used photocatalysts and analyze how specific structural changes alter their physicochemical and redox properties.


The Single-Cell Lab or How to Perform Single-Cell Molecular Analysis.
The increasing interest towards cellular heterogeneity within cell populations has pushed the development of new protocols to isolate and analyze single cells. PCR-based amplification techniques are widely used in this field. However, setting up an experiment and analyzing the results can sometimes be challenging. The aim of this chapter is to provide a general overview on single-cell PCR analysis focusing on the potential pitfalls and on the possible solutions to successfully perform the analysis.


Reinforcing cerclage in the prevention of preterm birth in women at high risk: a retrospective case-controlled study.
Transvaginal ultrasound measurement of cervical length is useful after suture insertion in predicting preterm delivery. However, there is little evidence to guide practice in the clinical scenario when fetal membranes are seen on ultrasound to be prolapsing distal to a cervical suture. To determine whether a reinforcing cerclage reduced preterm delivery in those women with ultrasound evidence of fetal membranes prolapsing distal to the first suture. A retrospective cohort study was conducted on women with a cervical suture in situ plus ultrasound evidence of fetal membranes prolapsing through the first suture. Exposed patients were those managed with a reinforcing cerclage. The unexposed group were women who were managed expectantly, without a reinforcing cerclage. Those women with a reinforcing cerclage were significantly more likely to deliver at an earlier gestation compared with those managed expectantly: 26(+0) (±5(+1) ) compared with 31(+1) (±7(+0) ) weeks, P = 0.047. More women in the reinforcing cerclage group delivered at <32 completed weeks' gestation: 12/13 (92%) versus 5/12 (42%), P = 0.01. There was no significant difference in the rate of second-trimester miscarriages between the expectant management group and those with a reinforcing cerclage: 2/12 (17%) versus 5/13 (38%), P = 0.38. Our study found that a reinforcing cerclage following primary cerclage failure hastened preterm delivery. The role of transvaginal ultrasound measurement of cervical length postsuture is debatable if the possible intervention is not beneficial and may be detrimental.


Fast-growing cyanobacteria bio-embedded into bacterial cellulose for toxic metal bioremediation.
Cyanobacterial biomass and cellulose-based materials have been used separately as green bio-adsorbents for the removal of toxic metals from water. Hybrid materials made of living microbial cells encased in a solid matrix have shown good potential for bioremediation. Here, the fast-growing cyanobacterium Synechococcus elongatus UTEX 2973 was embedded in situ into bacterial cellulose (BC), a robust biopolymer rich in hydroxyl groups with excellent water holding capacity. The living material was obtained by injecting S. elongatus into a Komagataeibacter sucrofermentans culture producing BC. Several types of BC/S. elongatus (BC/SE) materials were developed including small spheroids and flat films with different cyanobacteria loads via simple adjustments of the biosynthesis process parameters. BC/SE spheroids were evaluated for toxic copper removal and exhibited excellent adsorption properties compared to pure BC with a maximum capacity of 156.25 mg g-1. Thus, this simple bio-embedding approach holds promises in the development of living materials for environmental applications.


Brain plasticity after corpus callosum transection in the newborn rat.
The importance of afferent fibers in the final shaping of the central nervous system has been well established in several papers. In order to study how much one can influence cortical development by changing afferent input to the cortex cerebri, we transected corpus callosum in rats at 2 or 3 postnatal days. Equal numbers of lesioned and control animals were transcardially perfused with mixed aldehydes at postnatal day 6 (P-6) or P-14. Brain tissue was processed for Golgi impregnation, Nissl stain, and quantitative electron microscopic analysis. Morphometric image analysis (MOP 3) of the primary sensory area did not show significant differences with respect to cortical depth and total and apical dendritic length. The cross-sectional area and maximal diameter of neuronal cell bodies were smaller in P-14-lesioned animals. Ultrastructural analysis of the cortex revealed that synaptic density was not reduced in lesioned animals. In conclusion, early in development, rat brain can compensate for major changes in the commissural afferent system.


Regional distribution of acid phosphatase-positive axonal systems in the rat spinal cord and medulla, representing central terminals of cutaneous and visceral nociceptive neurons.
In addition to the substantia gelatinosa Rolandi, acid phosphatase active axonal systems are described (1) in the viscerosensory nucleus of the vagus nerve, (2) in Lissauer's band, (3) in the fasciculus cornus posterioris (Cajal), and (4) in the nucleus basilaris externus (Cajal). Electron microscopically, acid phosphatase is located in between synaptic vesicles of axon terminals; the vesicle population of such terminals in the Rolando substance, however, markedly differs from that in systems 1--4, characterized by the presence of large dense-core vesicles. While acid phosphatase-active axon terminals in the Rolando substance appear to subserve cutaneous nociception, circumstantial evidence suggests participation of systems 1--4 in processing visceral nociception.


Four-angle method for practical ultra-high-resolution magnetic resonance mapping of brain longitudinal relaxation time and apparent proton density.
Changes in longitudinal relaxation time (T1) and proton density (PD) are sensitive indicators of microstructural alterations associated with various central nervous system diseases as well as brain maturation and aging. In this work, we introduce a new approach for rapid and accurate high-resolution (HR) or ultra HR (UHR) mapping of T1 and apparent PD (APD) of the brain with correction of radiofrequency field, B1, inhomogeneities. The four-angle method (FAM) uses four spoiled-gradient recalled-echo (SPGR) images acquired at different flip angles (FA) and short repetition times (TRs). The first two SPGR images are acquired at low-spatial resolution and used to accurately map the active B1+ field with the recently introduced steady-state double angle method (SS-DAM). The estimated B1+ map is used in conjunction with the two other SPGR images, acquired at HR or UHR, to map T1 and APD. The method is evaluated with numerical, phantom, and in-vivo imaging measurements. Furthermore, we investigated imaging acceleration methods to further shorten the acquisition time. Our results indicate that FAM provides an accurate method for simultaneous HR or UHR mapping of T1 and APD in human brain in clinical high-field MRI. Derived parameter maps without B1+correction suffer from large inaccuracies, but this issue is well-corrected through use of the SS-DAM. Furthermore, the use of SPGR imaging with short TR and phased-array coil acquisition permits substantial imaging acceleration and enables robust HR or UHR T1 and APD mapping in a clinically acceptable time frame, with whole brain coverage obtained in less than 2 min or 5 min, respectively. The method exhibits high reproducibility and benefits from the use of the conventional SPGR sequence, available in all preclinical and clinical MRI machines, and very simple modeling to address a critical outstanding issue in neuroimaging.


Challenges for case-control studies with microbiome data.
In case-control studies of the human microbiome, the goal is to evaluate whether cases differ from controls in the microbiome composition of a particular body habitat and which taxa are responsible for the differences. These studies leverage sequencing technology and spectroscopy that provide new measurements of the microbiome. Three challenges in conducting reproducible microbiome research using a case-control design are compensating for differences in observed and actual microbial community composition, detecting "rare" taxa in microbial communities, and choosing properly powered analysis methods. The significance of each challenge, evaluation of commonly held views, analysis of unanswered questions, and suggestions of strategies for solutions are discussed. Understanding the effects of these choices on case-control analyses has been underappreciated, with an implicit assumption that further advances in technology will address all the current shortcomings. It is recommended that research on the human microbiome include positive and negative control experiments to provide insight into bias, contamination, and technical variation. Research protocols such as these may afford a better opportunity to make quantitative and qualitative adjustments to data, thereby reducing the risk of falsely positive results, increasing power to discover true disease determinants, and enhancing interpretation across studies.


Kindling in the posterior cingulate cortex: electrographic and behavioral characteristics.
Ictal, postictal and interictal electrical activities were recorded in the posterior cingulate cortex and hippocampus in the rat following kindling stimulations in the cingulate cortex. The afterdischarges (ADs) consisted of fast spikes and slow spike-and-wave of 2-5 Hz. Initial motor signs typically consisted of extension of the forelimbs and head, followed by clonic fore- and/or hind limb movements; there was no consistent laterality in any of the motor signs. Electrodes at or near the cingulum or corpus callosum ('white matter' group) gave kindling rates faster than other sites in the 'grey matter' of cingulate cortex (area 29); 2 of 7 animals had first-trial generalized motor seizures. The 'white matter' group averaged 4.6 ADs to reach the first 'bilateral forelimb clonus,' as compared to 17.8 ADs for the 'grey matter' group. The frequency and morphology of sharp EEG spikes in the cingulate cortex remained largely unchanged during the course of kindling. Cingulate interictal spikes were infrequent. The intimate connections of the cingulate cortex to motor and limbic structures rather than its intrinsic circuitry may be important for its fast kindling rates.


Effects of chronic lithium and sodium valproate on concentrations of brain amino acids.
The present study was designed to determine if the mood stabilizers, lithium and valproate, have common effects on concentrations of amino acid neurotransmitters which may be related to their mechanisms of action. Two separate groups of rats were administered therapeutic doses of lithium, sodium valproate, or saline for 2 weeks. Whole brain extracts were then examined using either high-field 1H NMR spectroscopy or HPLC. Both drugs decreased whole brain concentrations of aspartate, glutamate, and taurine while brain concentrations of gamma-aminobutyric acid (GABA) and alanine decreased following chronic sodium valproate administration but not following chronic lithium administration. These findings indicate that lithium and sodium valproate share common effects on the concentrations of certain amino acid neurotransmitters in whole brain which may be related to their mechanisms of action in bipolar disorder.


Development of customized implant and customized surgical osteotomy guide in ablative tumor surgery for accurate mandibular reconstruction.
The objective of this study was to provide the generalized methodology for design and development of a customized implant and customized surgical osteotomy guide (CSOG) for precise mandibular tumor resection and placement of a customized implant in ablative tumor surgery for accurate mandibular reconstruction. Medical imaging technique, image processing, virtual surgical planning (VSP), biomedical computer-aided design (CAD), and rapid prototyping (RP) were used to develop CSOG and customized implant. A mock surgical test and an experimental analysis were performed on the biomodel (RP assisted diseased model) to check the effectiveness of the CSOG. The paired t test showed the statistically significant result with the use of CSOG as compared to the without using CSOG in ablative mandibular tumor surgery. A mock test and an experimental analysis proved that, the precise tumor resection and customized implant placement with minimal gap between bone-implant junctions in mandibular reconstruction using CSOG.


Potent alpha-glucosidase inhibitors purified from the red alga Grateloupia elliptica.
Diabetes mellitus is a most serious and chronic disease whose incidence rates are increasing with incidences of obesity and aging of the general population over the world. One therapeutic approach for decreasing postprandial hyperglycemia is to retard absorption of glucose by inhibition of alpha-glucosidase. Two bromophenols, 2,4,6-tribromophenol and 2,4-dibromophenol, were purified from the red alga Grateloupia elliptica. IC(50) values of 2,4,6-tribromophenol and 2,4-dibromophenol were 60.3 and 110.4 microM against Saccharomyces cerevisiae alpha-glucosidase, and 130.3 and 230.3 microM against Bacillus stearothermophilus alpha-glucosidase, respectively. In addition, both mildly inhibited rat-intestinal sucrase (IC(50) of 4.2 and 3.6mM) and rat-intestinal maltase (IC(50) of 5.0 and 4.8mM). Therefore, bromophenols of G. elliptica have potential as natural nutraceuticals to prevent diabetes mellitus because of their high alpha-glucosidase inhibitory activity.


Impact of Dexmedetomidine Infusion on Opioid and Benzodiazepine Doses in Ventilated Pediatric Patients in the Cardiac Intensive Care Unit.
Dexmedetomidine (DEX) is frequently used as an adjunct agent for prolonged sedation in the intensive care unit (ICU), though its effect on concomitant opioids or benzodiazepines infusions is unclear. We explored the impact of DEX on concomitant analgosedation in a cohort of ventilated pediatric patients in a cardiac ICU, with stratification of patients according to duration of ventilation (< 5 versus ≥ 5 days) following DEX initiation. We conducted a retrospective analysis on ventilated patients receiving a DEX infusion ≥ 24 h and at least one other sedative/analgesic infusion (January 2011-June 2021). We evaluated trends of daily doses of opioids and benzodiazepines from 24 h before to 72 h following DEX initiation, stratifying patients based on ventilation duration after DEX initiation (< 5 versus ≥ 5 days). After excluding 1146 patients receiving DEX only, 1073 patients were included [median age 234 days (interquartile range 90, 879)]. DEX was associated with an opioid infusion in 99% of patients and a benzodiazepine infusion in 62%. Among patients ventilated for < 5 days (N = 761), opioids increased in the first 24 h following DEX initiation [+ 1.12 mg/kg/day (95% CI 0.96, 1.23), P < 0.001], then decreased [- 0.90 mg/kg/day (95% CI - 0.89, - 0.71), P < 0.001]; benzodiazepines slowly decreased [- 0.20 mg/kg/day (95% CI - 0.21, - 0.19), P < 0.001]. Among patients ventilated for ≥ 5 days (N = 312), opioid administration doubled [+ 2.09 mg/kg/day (95% CI 1.82, 2.36), P < 0.001] in the first 24 h, then diminished minimally [- 0.18 mg/kg/day (95% CI - 0.32, - 0.04), P = 0.015] without returning to baseline; benzodiazepine administration decreased minimally [- 0.03 mg/kg/day (95% CI - 0.05, - 0.01), P = 0.010]. Similar trends were confirmed when adjusting for age, gender, surgical complexity, recent major invasive procedures, duration of mechanical ventilation before DEX initiation, extubation within 72 h following DEX initiation, mean hourly DEX dose, and use of neuromuscular blocking infusion. While in patients ventilated < 5 days opioids initially increased and then quickly decreased in the 72 h following DEX initiation, among patients ventilated ≥ 5 days opioids doubled, then decreased only minimally; benzodiazepines decreased minimally in both groups, although more slowly in the long-ventilation cohort. These findings may inform decision-making on timing of DEX initiation in ventilated patients already being treated with opioid or benzodiazepine infusions.


Antiproliferative Activity of Some Newly Synthesized Substituted Nicotinamides Candidates Using Pyridine-2(1H) thione Derivatives as Synthon.
Some new pyridinethione and thienopyridine derivatives have been synthesized and evaluated for their antiproliferative activity using the MTT assay. Nicotinamide derivatives 3 have been synthesized and used for the preparation of new condensed thieno [2,3-b]pyridines by their reactions with active halo compounds. Finally the synthesized thienopyridine underwent ring closure whenever possible through boiling in a solution of sodium ethoxide. The antiproliferative evaluation against (HCT-116, HepG-2, and MCF-7) human cancer cells and one human healthy cell line (BJ-1) revealed that compounds 3b, 4c-5d, 7b-12a, 10d, and 13b have interesting antitumor activity specifically as antihepatocellular and anticolon cellular carcinoma agents. Besides, the docking results for most active derivatives were in agreement with the in vitro antitumor results.


Establishment of a human induced pluripotent stem cell line (CSUASOi008-A) from a type 2 diabetic patient with retinopathy.
Diabetic retinopathy (DR) is one of the most common and severe microvascular complications of diabetes, and the leading cause of preventable blindness in working-aged people. Here, we generated an induced pluripotent stem (iPS) cell line using blood-derived cells from a patient with DR. Peripheral blood mononuclear cells (PBMCs) were reprogrammed with Sendai virus.


Unintentional doping through the use of contaminated nutritional supplements.
To determine whether the intake of contaminated nutritional supplements could cause an athlete to fail a dope test. A contaminated nutritional supplement was used, identified in an ongoing study screening over-the-counter nutritional supplements. One capsule of the supplement, containing small amounts of 19-nor-4-androstenedione and 4-androsten-3,17-dione, not listed on the label, was administered to 5 healthy male volunteers. Fractional urine collection was done at prescribed intervals. Outcome measures. The samples were analysed using gas chromatography/mass spectrometry (GC/MS). Samples containing 19-norandrosterone, the main metabolite of 19-nor-4-androstenedione, were quantified using GC/MS. All the volunteers had urinary concentrations of 19-norandrosterone above the World Anti-Doping Agency threshold of 2 ng/ml from 2 hours post administration. In 2 volunteers 19-norandrosterone above the threshold value could still be detected beyond 36 hours post administration. The highest concentration of 19-norandrosterone found in a sample was 54.6 ng/ml at 8 hours post administration. The results of this study showed that the intake of microgram amounts of a prohibited substance in a nutritional supplement could cause an athlete to fail a dope test.


Prevalence of ponticulus posticus in a Northern Italian orthodontic population: a lateral cephalometric study.
The term "ponticulus posticus" refers to a partial or complete bony arch over the vertebral artery as it crosses the superolateral surface of the posterior arch of the atlas. This anatomical modification is linked to different symptoms, ranging from neckache to headache and migraine. This bony arch may also be incorrectly assessed during orthopaedic surgery for fixation of C1-C2, with consequent risk of damaging the vertebral artery. Its frequency in the general population has been widely analysed by literature in different geographic contexts, but an analysis of the prevalence of such feature in the Italian population is still missing. A Northern Italian orthodontic sample was analysed to assess the prevalence of ponticulus posticus. Lateral teleradiographies of 221 patients were examined. All the patients underwent lateral cephalometry for odontoiatric purposes and none of them was affected by congenital diseases or skeletal deformities. In the analysed sample, ponticulus posticus had a prevalence of 7.7 % for the complete form, and 9.0 % for the incomplete form. Complete and partial forms were observed, respectively, in 8.8 and 11.0 % of males, and in 6.9 and 7.7 % of females, without statistically significant differences. The current investigation provided the first data concerning the frequency of ponticulus posticus in Italy: further studies are needed to widen the sample, verify possible regional variations and improve the analysis by more advanced radiological examinations such as CT and cone beam CT scans.


The effects of prolonged ethanol exposure on the mechanical properties of polyurethane and silicone catheters used for intravascular access.
Products containing alcohol are commonly used with intravascular devices at insertion, to remove lipids from occluded intravascular devices used during parenteral nutrition, and increasingly for the prevention and treatment of intravascular device-related bloodstream infection. The effects of alcohol on the integrity of intravascular devices remain unknown. Two types of widely used commercial peripherally inserted central catheters, one made of polyetherurethane and one made of silicone, were exposed to a 70% ethanol lock solution for up to 10 weeks. Mechanical testing was performed to identify force-at-break, stress, strain, modulus of elasticity, modulus of toughness, and wall area of ethanol-exposed and control catheters. No significant differences between exposed and unexposed catheters were identified for any of the mechanical parameters tested except for a marginal reduction in the modulus of elasticity for both polyetherurethane and silicone catheters and minor increases in the wall area of polyetherurethane catheters. These data indicate that exposure to a 70% ethanol lock solution does not appreciably alter the integrity of selected commercial polyetherurethane and silicone catheters. Given the greatly expanded use of alcoholic solutions with intravascular devices of all types, we believe that manufacturers would be well advised to subject their catheters and other intravascular devices to formal testing of the type employed in this study.


Role of β2 adrenoreceptor gene polymorphism in the formation of cold hyperreactivity of the airways in asthmatics.
The relationship between β(2)-adrenoreceptor gene Arg16Gly polymorphism and bronchial cold reactivity has been studied. Genotype Arg16Arg and allele Arg16 carriership is associated with the development of cold bronchospasm in asthmatics. In addition, a significant reduction of 3,5'-cyclic adenosine monophosphate (cAMP) level has been recorded in Arg16Arg homozygotes on minute 30 after cold provocation in comparison with Gly16Gly genotype carriers. These data indicate the influence of primary dysfunction of β(2)-adrenoreceptors for the formation of bronchial cold hyperreactivity in the patients. Reduced cAMP synthesis by cells in Arg16Arg carriers indicates congenital liability of their β(2)-adrenoreceptors to desensitization during cold air isocapnic hyperventilation test.


SOHO State of the Art Updates and Next Questions: Updates on BTK Inhibitors for the Treatment of Chronic Lymphocytic Leukemia.
Over the last decade, targeted inhibition of Bruton's tyrosine kinase (BTK) has led to a paradigm shift in the way chronic lymphocytic leukemia (CLL) is managed. BTK inhibitors (BTKi) are broadly classified as covalent BTKI and noncovalent BTKi (cBTKi and ncBTK) Ibrutinib, as the first approved cBTKi, vastly improved outcomes for patients with CLL over prior chemoimmunotherapy regimens. However, long-term use is limited by both intolerance and resistance. The second generation of more selective BTKi were developed to improve tolerability. While these agents have led to an improved safety profile in comparison to Ibrutinib (both acalabrutinib and zanubrutinib), and improved efficacy (zanubrutinib), intolerance occasionally occurs, and resistance remains a challenge. The third generation of BTKi, which noncovalently or reversibly inhibits BTK, has shown promising results in early phase trials and are being evaluated in the phase 3 setting. These drugs could be an effective treatment option in patients with either resistance and intolerance to cBTKi. The most recent development in therapeutic agents targeting BTK is the development of BTK degraders. By removing BTK, as opposed to inhibiting it, these drugs could remain efficacious irrespective of BTK resistance mutations, however clinical data are limited at this time. This review summarizes the evolution and ongoing development of newer BTKi and BTK degraders in the management of CLL, with a focus of future directions in this field, including how emerging clinical data could inform therapeutic sequencing in CLL management.


Pediatric temporomandibular joint ankylosis and arthritis: Forgotten complications of acute otitis media.
Temporomandibular joint (TMJ) arthritis and ankylosis represent unusual but potential complications of ear suppuration, especially in children. We performed a review of the literature of pediatric otogenic TMJ arthritis and ankylosis, discussing their clinical and radiological features, their mechanism of infection spread, and the importance of a prompt diagnosis and treatment. We additionally describe a case of TMJ ankylosis following acute mastoiditis in a 4-year-old female patient. A search of English literature from January 1, 1980 to December 31, 2021 was performed on the electronic databases (PubMed, Web of Science and Scopus) in order to identify studies concerning TMJ complication after ear suppuration. Seventeen articles were considered eligible for the review. Eight and nine studies described otogenic TMJ ankylosis and arthritis, respectively. A total of 17 children affected by ankylosis consequent to ear infection and a total of 31 cases of TMJ arthritis concurrent to otomastoiditis were identified. Mean time elapsed between ear infection and diagnosis of TMJ ankylosis was 4.8 years (range 0.5-13). TMJ involvement during complicated otitis media should be kept in mind. Its prompt recognition is mandatory to set up appropriate treatment and follow-up and reduce the risk of ankylosis with its functional and psychological complications.


The Effect of Body Mass Index on Fluoroscopic Time and Radiation Dose During Sacroiliac Joint Injections.
Sacroiliac joint (SIJ) injections are commonly performed under fluoroscopic guidance. Radiation exposure to patients and providers has not been comprehensively studied, particularly the effect of body mass index (BMI). To identify whether patients with a larger BMI require longer fluoroscopy time or a larger radiation dose during performance of an SIJ injection. Retrospective study of an academic institution database. Academic outpatient musculoskeletal clinic. All patients who underwent SIJ injections during a 10-year period. Machine-reported fluoroscopic time and machine-reported radiation dose. A Bonferroni correction was implemented with P ≤ .01 as statistically significant. A total of 453 SIJ injections were performed in 359 patients. No statistically significant differences in fluoroscopy time were found between patients with BMI scores identified as normal, overweight, and obese (P = .054). However, the radiation doses were significantly greater for patients with higher BMI scores (χ(2) [2, n = 441] = 62.4, P < .001); the median (interquartile range) doses were 1210 (839), 1671 (1240), and 2090 (2170) mGy-cm(2) for normal weight, overweight, and obese patients, respectively. Although longer needles were used more often in obese patients (χ(2) [2, n = 452] = 31.5, P < .001), fluoroscopy time was not associated with needle length (P = .162). No relationships were identified between fluoroscopy time and first-time (as opposed to repeat) injection (P = .123), trainee involvement (χ(2) [1, n = 698] = 3.9, P = .049), or age (P = .337). Patients with an elevated BMI score who are undergoing SIJ injection receive an increased radiation dose despite equivalent fluoroscopic time. This finding suggests that the increased dose is likely due to x-ray output from the fluoroscope traversing a greater tissue mass, as opposed to the physician requiring more fluoroscopic images for proper needle placement. Fortunately, the increased radiation dose delivered to patients with a larger BMI score likely has negligible effects.


Epitope-based sieve analysis of Plasmodium falciparum sequences from a FMP2.1/AS02A vaccine trial is consistent with differential vaccine efficacy against immunologically relevant AMA1 variants.
To prevent premature dismissal of promising vaccine programs, it is critical to determine if lack of efficacy in the field is due to allele specific-efficacy, rather than to the lack of immunogenicity of the candidate antigen. Here we use samples collected during a field trial of the AMA1-based FMP2.1/AS02A malaria vaccine, which incorporates the AMA1 variant encoded by the reference Plasmodium falciparum 3D7 strain, to assess the usefulness of epitope-based sieve analysis for the detection of vaccine-induced allele-specific immune responses. The samples used are from volunteers who received the malaria vaccine FMP2.1/AS02A or a control (rabies vaccine), during a vaccine efficacy field trial, and who later developed malaria. In a previous study, P. falciparum DNA was extracted from all samples, and the ama1 locus amplified and sequenced. Here, a sieve analysis was used to measure T and B-cell escape, and difference in 3D7-like epitopes in the two treatment arms. Overall, no difference was observed in mean amino acid distance to the 3D7 AMA1 variant between sequences from vaccinees and controls in B-cell epitopes. However, we found a significantly greater proportion of 3D7-like T-cell epitopes that map to the AMA1 cluster one loop (c1L) region in the control vs. the vaccinee group (p = 0.02), consistent with allele-specific vaccine efficacy. Interestingly, AMA1 epitopes in infections from vaccinees had higher mean IC50, and consequently lower binding affinity, than epitopes generated from the control group (p = 0.01), suggesting that vaccine-induced selection impacted the immunological profile of the strains that pass through the sieve imposed by the vaccine-induced protection. These findings are consistent with a vaccine-derived sieve effect on the c1L region of AMA1 and suggest that sieve analyses of malaria vaccine trial samples targeted to epitopes identified in silico can help identify protective malaria antigens that may be efficacious if combined in a multivalent vaccine.


Predicting benefit from anti-angiogenic agents in malignancy.
A high probability of benefit is desirable to justify the choice of anti-angiogenic therapy from an ever-expanding list of expensive new anticancer agents. However, biomarkers of response to cytotoxic agents are not optimal for predicting benefit from anti-angiogenic drugs. This discussion will focus on both preclinical and clinical research to identify biomarkers for anti-angiogenic therapies that can inform dosing, early clinical benefit, initial drug choice, emerging resistance and second-line treatments.