Four classes of cell-associated proteoglycans in suspension cultures of articular-cartilage chondrocytes.
The characteristics of cell-associated proteoglycans were studied and compared with those from the medium in suspension cultures of calf articular-cartilage chondrocytes. By including hyaluronic acid or proteoglycan in the medium during [35S]sulphate labelling the proportion of cell-surface-associated proteoglycans could be decreased from 34% to about 15% of all incorporated label. A pulse-chase experiment indicated that this decrease was probably due to blocking of the reassociation with the cells of proteoglycans exported to the medium. Three peaks of [35S]sulphate-labelled proteoglycans from cell extracts and two from the medium were isolated by gel chromatography on Sephacryl S-500. These were characterized by agarose/polyacrylamide-gel electrophoresis, by SDS/polyacrylamide-gel electrophoresis of core proteins, by glycosaminoglycan composition and chain size as well as by distribution of glycosaminoglycans in proteolytic fragments. The results showed that associated with the cells were (a) large proteoglycans, typical for cartilage, apparently bound to hyaluronic acid at the cell surface, (b) an intermediate-size proteoglycan with chondroitin sulphate side chains (this proteoglycan, which had a large core protein, was only found associated with the cells and is apparently not related to the large proteoglycans), (c) a small proteoglycan with dermatan sulphate side chains with a low degree of epimerization, and (d) a somewhat smaller proteoglycan containing heparan sulphate side chains. The medium contained a large aggregating proteoglycan of similar nature to the large cell-associated proteoglycan and small proteoglycans with dermatan sulphate side chains with a higher degree of epimerization than those of the cells, i.e. containing some 20% iduronic acid.


High Performance Aqueous Li-Ion Flow Capacitor Realized Through Microstructure Design of Suspension Electrode.
Suspension electrode is the core of flowable electrochemical energy storage systems, which are considered suitable for large-scale energy storage. Nevertheless, obtaining suspension electrodes with both low viscosity and high conductivity is still a big challenge. In present work, spinel LiMn2O4 was chosen as an example to make suspension with low viscosity and high conductivity through microstructure morphology control of solid particles and the contact mode between active materials and conductive additives in suspension electrode. By coating a thin layer of polyaniline on the surface of spherical spinel LiMn2O4, the resulting suspension showed much higher electronic conductivity (about 10 times) and lower viscosity (about 4.5 times) as compared to irregular and bare spinel LiMn2O4-based suspension counterpart. As a result, the Li-ion flow capacitor based on LiMn2O4 and activated carbon suspensions exhibited a record energy density of 27.4 W h L-1 at a power density of 22.5 W L-1 under static condition to date, and can be smoothly work under an intermittent-flow mode. The strategy reported in this work is an effective way for obtaining suspension electrodes with low viscosity and high electronic conductivity simultaneously. It can not only be used in the flow capacitors, but also can be extended to other flowable electrochemical energy storage systems.


Functional expression of a Kir2.1-like inwardly rectifying potassium channel in mouse mammary secretory cells.
K(+) channels in mammary secretory (MS) cells are believed to play a role in transcellular electrolyte transport and thus determining ionic composition of the aqueous phase of milk. However, direct evidence for specific K(+) channel activity in native MS cells is lacking at the single-cell level. Here, we show for the first time that an inwardly rectifying K(+) (Kir) channel is functionally expressed in fully differentiated MS cells that were freshly isolated from the mammary gland of lactating mice. Using the standard whole cell patch-clamp technique, we found that mouse MS cells consistently displayed a K(+) current, whose electrophysiological properties are similar to those previously reported for Kir2.x channels, particularly Kir2.1: 1) current-voltage relationship with strong inward rectification, 2) slope conductance approximately proportional to the square root of external K(+) concentration, 3) voltage- and time-dependent and high-affinity block by external Ba(2+), and 4) voltage-dependent inhibition by external Cs(+). Accordingly, RT-PCR analysis revealed the gene expression of Kir2.1, but not Kir2.2, Kir2.3, and Kir2.4, in lactating mouse mammary gland, and immunohistochemical staining showed Kir2.1 protein expression in the secretory cells. Cell-attached patch recordings from MS cells revealed that a 31-pS K(+) channel with strong inward rectification was likely active at the resting membrane potential. Collectively, the present work demonstrates that a functional Kir2.1-like channel is expressed in lactating mouse MS cells. We propose that the channel might be involved, at least in part, in secretion and/or preservation of ionic components of milk stored into the lumen of these cells.


Parallel artificial liquid membrane extraction coupled with UPLC-ESI-MS/MS method for high-throughput quantitation of repaglinide in diabetic patients.
A high-throughput therapeutic monitoring method was developed for repaglinide (RPG) in diabetic patients, combining parallel artificial liquid membrane extraction (PALME) with ultraperformance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). PALME was performed using a 96-well donor plate comprising a donor solution containing a plasma sample, 50 mM phosphate buffer (pH = 8.0), and cetirizine (CTZ) as internal standard. A polypropylene (PP) porous membrane served as a selective support for the liquid membrane (SLM), preventing nonspecific binding produced by other membranes. The extraction was accomplished across SLM made of PP membrane with dodecyl acetate and 1 % trioctylamine (w/w), and the acceptor solution comprised DMSO and 200 mM formic acid (50:50, v/v). The simple workflow for PALME provided analyte enrichment, highly efficient sample cleanup, high throughput analysis, and excellent reproducibility. Method validation met FDA criteria, with a linear plasma calibration range (0.1-100 ng mL-1, r = 0.9995) and a lower limit of quantitation (LLOQ) of 0.1 ng mL-1. Recovery results at 98.9 % affirmed method reliability. The ability to analyze 198 samples per hour, coupled with a reduced amount of solvents, underscores the method's high throughput and eco-friendly profile. The PALME-UPLC-ESI-MS/MS method was successfully applied to therapeutic drug monitoring of RPG in diabetic patients following 2 mg RPG tablet administration, establishing its effectiveness.


Risk factors, pregnancy complications, and prevention of hypertensive disorders in women with pregravid diabetes mellitus.
To review the rate, risk factors, and potential prevention of preeclampsia in women with pregravid diabetes mellitus. Detailed review of recent English literature describing pregnancy outcome in women with pregravid insulin-dependent diabetes mellitus (Type 1 diabetes). The primary focus of the review is risk factors for preeclampsia and potential methods suggested for prevention of preeclampsia. In addition, pitfalls in diagnosis of preeclampsia will be emphasized. The reported rate of preeclampsia ranged from 9% to 66%. This rate increased with increased severity of diabetes by White's classification, with the highest rate reported in women with pregravid diabetic nephropathy. Risk factors identified for preeclampsia were nulliparity, chronic hypertension, microalbuminuria prior to pregnancy, nephropathy, and poor glycemic control early in pregnancy. There is lack of agreement among reports in the literature concerning criteria used to diagnose preeclampsia. There are two randomized trials that included women with Type 1 diabetes who were randomized to receive either low-dose aspirin or placebo. There was no reduction in the rate of preeclampsia with low-dose aspirin in either trial. In women with pregravid Type 1 diabetes, the rates of preeclampsia and adverse neonatal outcome increase with increased severity of diabetes. Low-dose aspirin does not reduce rate of preeclampsia in such women. Further studies should address the role of good glycemic control prior to 20 weeks gestation and prevention of microalbuminuria.


Facile one-pot synthesis of a aptamer-based organic-silica hybrid monolithic capillary column by "thiol-ene" click chemistry for detection of enantiomers of chemotherapeutic anthracyclines.
In the current study, we developed a facile strategy for the one-pot synthesis of an aptamer-based organic-silica hybrid monolithic capillary column. A 5'-SH-modified aptamer, specifically targeting doxorubicin, was covalently modified in the hybrid silica monolithic column by a sol-gel method combined with "thiol-ene" click reaction. The prepared monolithic column had good stability and permeability, large specific surface, and showed excellent selectivity towards chemotherapeutic anthracyclines of doxorubicin and epirubicin. In addition, the enantiomers of doxorubicin and epirubicin can be easily separated by aptamer-based affinity monolithic capillary liquid chromatography. Furthermore, doxorubicin and epirubicin spiked in serum and urine were also successfully determined, which suggested that the complex biological matrix had a negligible effect on the detection of doxorubicin and epirubicin. Finally, we quantified the concentration of epirubicin in the serum of breast cancer patients treated with epirubicin by intravenous injection. The developed analytical method is cost-effective and rapid, and biological samples can be directly analyzed without any tedious sample pretreatment, which is extremely useful for monitoring medicines in serum and urine for pharmacokinetic studies.


Postsynthetic ionization of an imidazole-containing metal-organic framework for the cycloaddition of carbon dioxide and epoxides.
A bifunctional imidazolium functionalized zirconium metal-organic framework (Zr-MOF), (I-)Meim-UiO-66 (2), was successfully prepared from the imidazole-containing Zr-MOF Im-UiO-66 (1) by a post-synthetic modification (PSM) method. It was found that the crystal size and pore features of the imidazole-containing 1 could be tuned at the nanoscale. The bifunctional MOF 2, containing Brønsted acid sites and iodide ions, was shown to be an efficient and recyclable heterogeneous catalyst for the cycloaddition of carbon dioxide (CO2) with epoxides, without the use of any co-catalyst, at ambient pressure. The solvent-free synthesis of the cyclic carbonate from CO2 and an epoxide was monitored by in situ Fourier transform infrared spectroscopy (FT-IR) and an acid/base synergistic catalysis mechanism was proposed. We hope that our strategy provides an effective approach for the introduction of functional N-heterocyclic groups into MOFs for potential applications.


Discordance in TKA expectations between patients and surgeons.
Aligning patient and surgeon expectations preoperatively may lead to better postoperative medical and rehabilitation compliance and therefore improve outcomes and increase satisfaction. We (1) determined the rate of discordantly high patient expectations compared with those of their surgeon in patients undergoing TKA; and (2) evaluated the impact of the preoperative educational class, patient characteristics, and functional status on the likelihood of having discordantly high patient expectations. We enrolled 205 patients awaiting TKA. Each patient completed a validated questionnaire that addresses expectations of postoperative pain relief, function, and well-being as part of a preoperative assessment. The surgeon completed the same expectations questionnaire preoperatively blinded to their patient's response. Patients had discordantly high expectations if their scores were ≥ 7 points higher than the surgeon on a 0 to 100 score range. Regression analysis was performed to determine the effect of class, patient characteristics, and functional status on the likelihood of having discordantly high patient expectations. Thirty-seven percent of the patients had expectation scores ≥ 7 points higher than those of their surgeon. Patients were less likely to have discordantly higher expectations if they were female (OR, 0.56; CI, 0.32-0.97) and if their pain level was high (OR, 0.99; CI, 0.98-0.99). Patients were more likely to have discordantly higher expectations if they filled out the expectations survey before rather than after the preoperative educational class (OR, 1.80; CI, 1.08-3.01). With increasing TKA use, surgeons will likely encounter more patients with discordantly high expectations. The preoperative educational class can be used to target patients more likely to have discordantly high expectations.


[Pro-apoptotic function of hepatitis B virus X protein].
Infection of hepatitis B virus (HBV) is a main cause of liver diseases including hepatitis, cirrhosis and hepatocellular carcinoma (HCC). Among the HBV-encoded proteins, the HBV X protein (HBx) has been suspected to be strongly involved in HBV-associated liver pathogenesis. HBx, a virally encoded multifunctional regulator, has been shown to induce apoptosis, anti-apoptosis, proliferation, and transformation of cells depending on the cell lines, model systems used, assay protocols, and research groups. Among the several activities of HBx, the pro-apoptotic function of HBx will be discussed in this review. Given that the disruption of apoptosis pathway by HBx contributes to the liver pathogenesis, a better understanding of the molecular interference in the cellular pro-apoptotic networks by HBx will provide useful clues for the intervention in HBV-mediated liver diseases.


Microsatellites provide insight into contrasting mating patterns in arribada vs. non-arribada olive ridley sea turtle rookeries.
Molecular studies of sea turtles have shown that the frequency of multiple paternity (MP) varies between species, and between rookeries of the same species. This study uses nuclear microsatellite markers to compare the incidence of MP in two neighbouring olive ridley rookeries on the Pacific coast of Costa Rica, with contrasting nesting behaviours -- the 'arribada' population nesting at Ostional and the solitary nesters of Playa Hermosa. Using two highly polymorphic microsatellite markers, we tested 13 nests from each location and found a significant difference (P < 0.001) between the level of MP of the arribada rookery (92%- the highest found for marine turtles) and that of the solitary nesting rookery (30%). Additional analyses based on six microsatellite loci revealed no genetic differentiation between nesting females from the two locations, or between nesting females and attendant males from the Ostional breeding area. Sixty-nine per cent of the nests with MP were fathered by a minimum of three different males, and three nests showed evidence of at least four fathers. The results suggest that the differences observed in levels of MP between arribada and solitary rookeries are due to an effect of abundance of individuals on the mating system. This is supported by a regression analysis combining other paternity studies on sea turtles which shows that levels of MP increase with increasing abundance of nesting females.


Molecular Characteristics and Zoonotic Potential of Salmonella Weltevreden From Cultured Shrimp and Tilapia in Vietnam and China.
Salmonella Weltevreden is increasingly reported from aquatic environments, seafood, and patients in several Southeast Asian countries. Using genome-wide analysis, we characterized S. Weltevreden isolated from cultured shrimp and tilapia from Vietnam and China to study their genetic characteristics and relatedness to clinical isolates of S. Weltevreden ST-365. The phylogenetic analysis revealed up to 312 single-nucleotide polymorphism (SNP) difference between tilapia isolates, whereas isolates from shrimp were genetically more closely related. Epidemiologically unrelated isolates from Vietnam were closely related to isolates from China, e.g., 20 SNPs differences between strains 28V and 75C. In comparison with strains from other parts of the world, our environmental isolates predominantly clustered within the continental South Asia lineage, constituted mostly of strains from human stool with as low as seven SNPs difference, e.g., 30V versus Cont_ERR495254. All sequenced isolates were MLST type ST-365 and contained the major virulence-related genes encoded by the Salmonella Pathogenicity Islands 1-5. Ten of the isolates harbored the IncFII(S) plasmid similar to the virulence genes-mediated plasmid pSPCV of S. Paratyphi C, and one isolate had the IncQ1 plasmid on the same contig with strA/B, sul2, and tetA resistance genes similar to the IncQ1 type, pNUC of S. Typhimurium. A pangenomic analysis yielded 7891 genes including a core genome of 4892 genes, with a closely related accessory genome content between clinical and environmental isolates (Benjamini p > 0.05). In a search for differences that could explain the higher prevalence of S. Weltevreden in aquatic samples, genomes were compared with those of other Salmonella enterica serovars. S. Weltevreden revealed specific regions harboring glpX (Fructose-1;6-bisphosphatase; class II), rfbC (dTDP-4-dehydrorhamnose 3;5-epimerase), and cmtB (PTS Mannitol-specific cryptic phosphotransferase enzyme IIA component) involved in carbohydrate biosynthesis pathways. Our study builds grounds for future experiments to determine genes or pathways that are essential when S. Weltevreden are in aquatic environments and microbial interactions providing survival advantages to S. Weltevreden in such environments.


Ceramide recruits and activates protein kinase C zeta (PKC zeta) within structured membrane microdomains.
We have previously demonstrated that hexanoyl-D-erythro-sphingosine (C(6)-ceramide), an anti-mitogenic cell-permeable lipid metabolite, limited vascular smooth muscle growth by abrogating trauma-induced Akt activity in a stretch injury model of neointimal hyperplasia. Furthermore, ceramide selectively and directly activated protein kinase C zeta (PKC zeta) to suppress Akt-dependent mitogenesis. To further analyze the interaction between ceramide and PKC zeta, the ability of ceramide to localize within highly structured lipid microdomains (rafts) and activate PKC zeta was investigated. Using rat aorta vascular smooth muscle cells (A7r5), we now demonstrate that C(6)-ceramide treatment results in an increased localization and phosphorylation of PKC zeta within caveolin-enriched lipid microdomians to inactivate Akt. In addition, ceramide specifically reduced the association of PKC zeta with 14-3-3, a scaffold protein localized to less structured regions within membranes. Pharmacological disruption of highly structured lipid microdomains resulted in abrogation of ceramide-activated, PKC zeta-dependent Akt inactivation, whereas molecular strategies suggest that ceramide-dependent PKC zeta phosphorylation of Akt3 at Ser(34) was necessary for ceramide-induced vascular smooth muscle cell growth arrest. Taken together, these data demonstrate that structured membrane microdomains are necessary for ceramide-induced activation of PKC zeta and resultant diminished Akt activity, leading to vascular smooth muscle cell growth arrest.


Outcome of community-acquired pneumonia: influence of age, residence status and antimicrobial treatment.
Community-acquired pneumonia remains a major cause of mortality in developed countries. There is much discrepancy in the literature regarding factors influencing the outcome in the elderly population. Data were derived from a multicentre prospective study initiated by the German Competence Network for Community-Acquired Pneumonia. Patients with community-acquired pneumonia (n = 2,647; 1,298 aged < 65 yrs and 1,349 aged > or = 65 yrs) were evaluated, of whom 72.3% were hospitalised and 27.7% treated in the community. Clinical history, residence status, course of disease and antimicrobial treatment were prospectively documented. Microbiological investigations included cultures and PCR of respiratory samples and blood cultures. Factors related to mortality were included in multivariate analyses. The overall 30-day mortality was 6.3%. Elderly patients exhibited a significantly higher mortality rate that was independently associated with the following: age; residence status; confusion, urea, respiratory frequency and blood pressure (CURB) score; comorbid conditions; and failure of initial therapy. Increasing age remained predictive of death in the elderly. Nursing home residents showed a four-fold increased mortality rate and an increased rate of gram-negative bacillary infections compared with patients dwelling in the community. The CURB score and cerebrovascular disease were confirmed as independent predictors of death in this subgroup. Age and residence status are independent risk factors for mortality after controlling for comorbid conditions and disease severity. Failure of initial therapy was the only modifiable prognostic factor.


What Predicts Improvement of Dizziness after Multimodal and Interdisciplinary Day Care Treatment?
Vertigo and dizziness are common in community-dwelling people and can be treated in specialized multidisciplinary settings. To develop tailored interventions, however, we have to explore risk factors for favorable and unfavorable outcomes. We prospectively investigated patients with chronic vertigo and dizziness subjected to our 5-day multimodal and interdisciplinary day care treatment in the Center for Vertigo and Dizziness of Jena University Hospital, Germany. The Vertigo Severity Scale (VSS), the Body Sensations Questionnaire (BSQ), the Hospital Anxiety and Depression Scale (HADS), the Agoraphobic Cognitions Questionnaire (ACQ), the Mobility Inventory (MI), and the burden and intensity of dizziness (using a visual analogue scale) were assessed at baseline (n = 754) and after 6 months (n = 444). In addition, 14 Likert-scaled questions were used to quantify the change in personal attitude and behavior towards the complaints after 6 months. Dizziness-related burden and intensity improved with a large effect size. The largest improvement was seen in the attitudes towards dizziness, the understanding of somatic causes, and the perceived ability to influence dizziness. However, the ability to work and to carry out professional activity was improved to a lesser extent. The overall improvement of dizziness was associated with the absence of a depressive mood, a short duration of vertigo, a lower VSS, a lower perceived intensity of vertigo, and distinct vertigo diagnoses, namely Meniere's disease, vestibular migraine, vestibular neuritis, vestibular paroxysmia, and vestibular schwannoma. Worsening of dizziness/vertigo was associated with depressive symptoms, permanent vertigo, distinct vertigo diagnoses (central vertigo, multisensory deficit), and a higher perceived burden due to vertigo. The six-month outcome of patients with dizziness presented to a specialized outpatient clinic appears to be favorable. Nevertheless, people with the abovementioned risk factors at baseline have less benefit and probably need adapted and tailored vertigo interventions to improve long-term outcome.


Mentors' perceptions and experiences of supporting student nurses in practice.
This paper reports on a funded project that explored the perceptions and experiences of mentors regarding student nurse support in practice. The study employed a mixed-method approach, using questionnaires and focus groups with mentors from one acute Trust and one community Trust. The findings highlighted the multifaceted nature of student learning in practice, with mentors reporting that clinical skills, adjustment to the placement and integrating into the team were the aspects students needed most support with. Mentors were aware of their roles and responsibilities in supporting students and recognized the importance of their own personal attributes. The participants reported a number of challenges, particularly time, competing demands and paperwork, and suggested that a team approach and support groups could help to overcome these. The support for students provided by peers and health-care assistants was recognized, as was the need to ensure that students are prepared to take responsibility for their learning.


Bacterial toxins: Offensive, defensive, or something else altogether?
The secretion of proteins that damage host tissue is well established as integral to the infectious processes of many bacterial pathogens. However, recent advances in our understanding of the activity of toxins suggest that the attributes we have assigned to them from early in vitro experimentation have misled us into thinking of them as merely destructive tools. Here, we will discuss the multifarious ways in which toxins contribute to the lifestyle of bacteria and, by considering their activity from an evolutionary perspective, demonstrate how this extends far beyond their ability to destroy host tissue.


Global impact of antibacterial resistance in patients with hematologic malignancies and hematopoietic cell transplant recipients.
Patients with hematologic malignancies and hematopoietic cell transplant (HCT) recipients are at high risk of developing bacterial infections. These patients may suffer severe consequences from these infections if they do not receive immediate effective therapies, and thus are uniquely threatened by antimicrobial-resistant bacteria. Here, we outline how the emergence of specific resistant bacteria threatens the effectiveness of established approaches to prevent and treat infections in this population. The emergence of fluoroquinolone resistance among Enterobacterales and viridans group streptococci may decrease the effectiveness of fluoroquinolone prophylaxis during neutropenia. The emergence of Enterobacterales that produce extended-spectrum β-lactamases or carbapenemases and of increasingly resistant Pseudomonas aeruginosa may result in neutropenic patients experiencing delayed time to active antibacterial therapy, and consequently worse clinical outcomes. The ability to select targeted antibacterial therapies after the availability of susceptibility data may be limited in patients infected with metallo-β-lactamase-producing Enterobacterales and difficult-to-treat P. aeruginosa. Vancomycin-resistant enterococci and Stenotrophomonas maltophilia can cause breakthrough infections in patients already being treated with broad-spectrum β-lactam antibiotics. Resistance can also limit the ability to provide oral stepdown antibacterial therapy for patients who could otherwise be discharged from hospitalization. We also outline strategies that have the potential to mitigate the negative impact of antimicrobial resistance, including interventions based on active screening for colonization with resistant bacteria and the use of novel rapid diagnostic assays. Additional research is needed to better understand how these strategies can be leveraged to combat the emerging crisis of antimicrobial resistance in patients with hematologic malignancies and HCT recipients.


Bacterial Metabolism in the Host Environment: Pathogen Growth and Nutrient Assimilation in the Mammalian Upper Respiratory Tract.
Pathogens evolve in specific host niches and microenvironments that provide the physical and nutritional requirements conducive to their growth. In addition to using the host as a source of food, bacterial pathogens must avoid the immune response to their presence. The mammalian upper respiratory tract is a site that is exposed to the external environment, and is readily colonized by bacteria that live as resident flora or as pathogens. These bacteria can remain localized, descend to the lower respiratory tract, or traverse the epithelium to disseminate throughout the body. By virtue of their successful colonization of the respiratory epithelium, these bacteria obtain the nutrients needed for growth, either directly from host resources or from other microbes. This chapter describes the upper respiratory tract environment, including its tissue and mucosal structure, prokaryotic biota, and biochemical composition that would support microbial life. Neisseria meningitidis and the Bordetella species are discussed as examples of bacteria that have no known external reservoirs but have evolved to obligately colonize the mammalian upper respiratory tract.


Explanation of [Formula: see text] fractional quantum Hall state in bilayer graphene.
The commensurability condition is applied to determine the hierarchy of fractional filling of Landau levels for fractional quantum Hall effect (FQHE) in monolayer and bilayer graphene. Good agreement with experimental data is achieved. The presence of even-denominator filling fractions in the hierarchy of the FQHE in bilayer graphene is explained, including the state at [Formula: see text].


Experienced operators achieve superior patency and wound complication rates with endoscopic great saphenous vein harvest compared with open harvest in lower extremity bypasses.
Prior studies have suggested improved wound complication rates but decreased primary patency in lower extremity bypasses performed with endoscopic vein harvest (EVH) vs open vein harvest (OVH). We hypothesize that the inferior patency reflects the initial learning curve for EVH and that improved patency can be achieved with experience. This was a single-institution review of 113 patients with critical limb ischemia who underwent infrainguinal bypass with a continuous segment of great saphenous vein harvested endoscopically (n = 49) or through a single open incision (n = 64) from 2012 to 2017. EVH was performed by surgeons with >5 years' experience with this technique. Operative outcomes, patency, complications, and readmission rates were compared between the harvest methods. EVH data were also compared with our prior reported series of our initial experience with this technique to determine the effects of experience on outcomes. There were no significant differences in patient demographics, medications, operative indications, or inflow/outflow vessels between the two groups. Mean operative time was 322 minutes and median hospital length of stay was 6 days for OVH, and was 340 minutes and 5 days for EVH, which was not significant. Harvest-related wound complications were more frequent with OVH (28% vs 2%, P < .001). Primary patency at 1 and 3 years was 65% and 58% for OVH, and 79% and 71% for EVH, respectively (P = .18), assisted primary patency was 77% and 74% for OVH and 94% and 89% for EVH, respectively (P = .05), and secondary patency was 82% and 79% for OVH and 95% and 95% for EVH, respectively (P = .03). The 30-day readmission rates were similar between OVH (20%) and EVH (12%, P = .26), but 90-day readmissions were more frequent in the OVH group (34% vs 14%, P = .018). Compared with our earlier series of EVH, the current cohort had significantly improved 3-year primary (71% vs 42%, P = .012), primary assisted patency (89 vs 66%, P = .034), and secondary patency (95% vs 66%, P = .003). With experience, lower extremity bypass using EVH can result in improved patency compared with OVH and initial EVH use, while also resulting in fewer wound complications and readmissions, with comparable operative times and hospital length of stay. This technique should be more widely adopted by vascular surgeons as a primary method of vein harvest.


B-N polymer embedded iron(0) nanoparticles as highly active and long lived catalyst in the dehydrogenation of ammonia borane.
B-N polymer embedded iron(0) nanoparticles (NPs) were in-situ generated from the reduction of iron(III) acetylacetonate during the dehydrogenation of ammonia borane (AB) in THF solution at 40.0 +/- 0.5 degrees C. The iron(0) NPs could be isolated as powder from the reaction solution by centrifugation and characterized by UV-Vis, TEM, and XRD. They are redispersible in polar solvent such as THF and yet highly active catalysts in the dehydrogenation of AB providing a TOF value of 202 h(-1) at 40.0 +/- 0.5 degrees C. The catalytic activity of iron(0) NPs compare well with those of the known homogenous and heterogeneous precious metal catalysts reported so far. They are also long-life catalysts in the dehydrogenation of AB providing 1410 turnovers over 18 h at 40.0 +/- 0.5 degrees C. The poisoning experiments using carbon disulfide show that the dehydrogenation of AB catalyzed by iron(0) NPs is a heterogeneous catalysis. The catalytic dehydrogenation of AB in the presence of iron(0) NPs was followed by measuring the volume of hydrogen generated and by 11B-NMR spectroscopy. Our report also includes the results of a detailed kinetic study on the catalytic dehydrogenation of AB depending on the catalyst concentration, substrate concentration, and temperature. The dehydrogenation of AB produces sparingly soluble B-N polymers which provide just enough stability to the iron(0) NPs. The co-precipitation of some iron(0) NPs with the sparingly soluble polymers causes a slight decrease in the catalytic activity toward the end of dehydrogenation. However, iron(0) NPs embedded in B-N polymers appear to be an efficient catalyst in hydrogen generation from ammonia borane at moderate temperature.


Consequences of measurement error in qPCR telomere data: A simulation study.
The qPCR method provides an inexpensive, rapid method for estimating relative telomere length across a set of biological samples. Like all laboratory methods, it involves some degree of measurement error. The estimation of relative telomere length is done subjecting the actual measurements made (the Cq values for telomere and a control gene) to non-linear transformations and combining them into a ratio (the TS ratio). Here, we use computer simulations, supported by mathematical analysis, to explore how errors in measurement affect qPCR estimates of relative telomere length, both in cross-sectional and longitudinal data. We show that errors introduced at the level of Cq values are magnified when the TS ratio is calculated. If the errors at the Cq level are normally distributed and independent of true telomere length, those in the TS ratio are positively skewed and proportional to true telomere length. The repeatability of the TS ratio declines sharply with increasing error in measurement of the Cq values for telomere and/or control gene. In simulated longitudinal data, measurement error alone can produce a pattern of low correlation between successive measures of relative telomere length, coupled with a strong negative dependency of the rate of change on initial relative telomere length. Our results illustrate the importance of reducing measurement error: a small increase in error in Cq values can have large consequences for the power and interpretability of qPCR estimates of relative telomere length. The findings also illustrate the importance of characterising the measurement error in each dataset-coefficients of variation are generally unhelpful, and researchers should report standard deviations of Cq values and/or repeatabilities of TS ratios-and allowing for the known effects of measurement error when interpreting patterns of TS ratio change over time.


[Etiological significance of rotaviruses in acute intestinal diseases in children].
The paper presents the results of virological, serological, and epidemiological examinations of patients with acute enteric infections of obscure etiology carried out for detection of rotavirus gastroenteritides. Rotaviruses have been found to play a certain role in generating acute gastroenteritides in infants under 3 years, mostly among infants, of 1-2 years, the least among those of 0-6 months of age. Adult family members were the source of infection for the children. The season of infection falls on October-December.


Structural and dynamic perturbations induced by heme binding in cytochrome b5.
The water-soluble domain of rat hepatic cytochrome b(5) undergoes marked structural changes upon heme removal. The solution structure of apocytochrome b(5) shows that the protein is partially folded in the absence of the heme group, exhibiting a stable module and a disordered heme-binding loop. The quality of the apoprotein structure in solution was improved with the use of heteronuclear NMR data. Backbone amide hydrogen exchange was studied to characterize cooperative units in the protein. It was found that this criterion distinguished the folded module from the heme-binding loop in the apoprotein, in contrast to the holoprotein. The osmolyte trimethylamine N-oxide (TMAO) did not affect the structure of the apoprotein in the disordered region. TMAO imparted a small stabilization consistent with an unfolded state effect correlating with the extent of buried surface area in the folded region of the native apoprotein. The failure of the osmolyte to cause large conformational shifts in the disordered loop supported the view that the specificity of the local sequence for the holoprotein fold was best developed with the stabilization of the native state through heme binding. To dissect the role of the heme prosthetic group in forcing the disordered region into the holoprotein conformation, the axial histidine belonging to the flexible loop (His63) was replaced with an alanine, and the structural properties of the protein with carbon-monoxide-ligated reduced iron were studied. The His63Ala substitution resulted in a protein with lower heme affinity but nevertheless capable of complete refolding. This indicated that the coordination bond was not necessary to establish the structural features of the holoprotein. In addition, the weak binding of the heme in this protein resulted in conformational shifts at a location distant from the binding site. The data suggested an uneven distribution of cooperative elements in the structure of the cytochrome.


Clinical outcome measures and rating scales in multiple sclerosis trials.
In this review, we analyzed clinical outcome measures used in multiple sclerosis (MS) clinical trials in which the primary goal is to slow or arrest progression of disease. In addition, we examined rating scales that quantify symptomatic complications of MS (for example, spasticity) and the current role of magnetic resonance imaging in MS treatment trials. Each proposed scale has advantages and deficiencies, and none meets all the criteria for an ideal outcome measure. The validity of trial design may be improved by using combinations of selected components of current scales as well as new instruments targeted to specific variables (such as motor strength). Symptom-specific rating scales are most appropriately used in trials of symptomatic therapeutic strategies for MS. Until serial magnetic resonance imaging changes are definitely known to predict long-term impairment and disability in patients with MS, clinical outcome measures will remain the primary means of assessing therapeutic efficacy in phase III clinical trials.


Avelumab Plus Axitinib as First-Line Therapy for Advanced Renal Cell Carcinoma: Long-Term Results from the JAVELIN Renal 100 Phase Ib Trial.
Progression-free survival was significantly longer in patients who received avelumab plus axitinib versus sunitinib as first-line treatment for advanced renal cell carcinoma (aRCC) in a randomized phase III trial. We report long-term safety and efficacy of avelumab plus axitinib as first-line treatment for patients with aRCC from the JAVELIN Renal 100 phase Ib trial (NCT02493751). In this open-label, multicenter, phase Ib study, patients with untreated aRCC received avelumab 10 mg/kg every 2 weeks plus axitinib 5 mg twice daily or with axitinib for 7 days followed by avelumab plus axitinib. Safety and efficacy were assessed in all patients receiving at least one dose of avelumab or axitinib. Overall, 55 patients were enrolled and treated. Median follow-up was 55.7 months (95% CI, 54.5-58.7). Treatment-related adverse events of any grade or grade ≥3 occurred in 54 (98.2%) and 34 (61.8%) patients, respectively. The confirmed objective response rate was 60.0% (95% CI, 45.9-73.0), including complete response in 10.9% of patients. Median duration of response was 35.9 months (95% CI, 12.7-52.9); the probability of response was 65.8% (95% CI, 46.7-79.4) at 2 years. Median progression-free survival was 8.3 months (95% CI, 5.3-32.0). Median overall survival was not reached (95% CI, 40.8-not estimable); the 5-year overall survival rate was 57.3% (95% CI, 41.2-70.5). Five-year follow-up for combination treatment with avelumab plus axitinib in previously untreated patients with aRCC showed long-term clinical activity with no new safety signals, supporting use of this regimen within its approved indication in clinical practice (Clinicaltrials.gov NCT02493751).


Vector population manipulation for control of arboviruses--a novel prospect for India.
India, the seventh largest country in the world, has diverse geographical and climatic regions with vast rural and peri-urban areas. Many are experiencing an escalation in the spread and intensity of numerous human diseases transmitted by insects. Classically, the management of these vector-borne diseases is underpinned by either chemical insecticides and/or environmental management targeted at the vector. However, these methods or their present implementation do not offer acceptable levels of control, and more effective and sustainable options are now available. Genetic strategies for the prevention of arbovirus transmission are most advanced for dengue and chikungunya, targeting their primary vector, Aedes aegypti. The national burden in terms of morbidity and mortality as a direct consequence of dengue virus in India is considered to be the largest worldwide, over 4 times that of any other country. Presently, new genetic technologies are undergoing field evaluation of their biosafety and efficacy in several countries. This paper discusses the merits of these approaches and argues for fair and transparent appraisal in India as a matter of urgency. Identification of any associated risks and their appropriate mitigation are fundamental to that process.


Hyperprolactinemia and possibly related development of prolactinoma during amisulpride treatment; three cases.
Schizophrenia is a chronic and debilitating psychotic mental disorder that affects about 1% of the world's population. Antipsychotic drugs are the mainstay of treatment in schizophrenia. Hyperprolactinemia, which is a common side effect of typical antipsychotics, is also associated with the use of some of the newer atypical agents. Antipsychotics may enhance prolactinoma growth as manifested by an increase in serum prolactin concentration. Prolactin-secreting pituitary adenomas possibly related with antipsychotics have been described in the literature. To our knowledge, this is the first series of cases showing a possible relation between pituitary adenomas and amisulpride treatment in patients with schizophrenia.


[A severe stroke following carotid sinus massage].
Carotid sinus massage is a widely used method for diagnosis and treatment of supraventricular tachycardia and carotid sinus hypersensitivity. Complications, mostly neurological, can occur but are rare. Carotid stenosis is a risk factor for complications. Hearing a carotid bruit on auscultation indicates stenosis, and is a contraindication for performing carotid sinus massage. However, the sensitivity of auscultation is insufficient. A 71-year-old man with a history of hypertension and hypercholesterolemia presented to the cardiology accident and emergency department with palpitations. A supraventricular tachycardia was found on examination, for which carotid sinus massage was performed. The patient developed severe aphasia and right-sided hemiparesis caused by an extensive stroke, and died a few days later. The chance of complications following carotid sinus massage is slight; however, this type of complication can have severe consequences. Safer alternative methods may be used for patients with supraventricular tachycardia. In older patients with vascular risk factors, more extensive diagnostic investigations for carotid stenosis should be considered in the diagnostic workup for carotid sinus hypersensitivity.


Elucidating mechanisms of solvent toxicity in ethanologenic Escherichia coli.
Ethanol toxicity and its effect on ethanol production by the recombinant ethanologenic Escherichia coli strain KO11 were investigated in batch and continuous fermentation. During batch growth, ethanol produced by KO11 reduced both the specific cell growth rate (micro) and the cell yield (Y(X/S)). The extent of inhibition increased with the production of both acetate and lactate. Subsequent accumulation of these metabolites and ethanol resulted in cessation of cell growth, redirection of metabolism to reduce ethanol production, and increased requirements for cell maintenance. These effects were found to depend on both the glycolytic flux and the flux from pyruvate to ethanol. Pyruvate decarboxylase (Pdc) and alcohol dehydrogenase (Adh) activities measured during the batch fermentation suggested that decreased ethanol production resulted from enzyme inhibition rather than down-regulation of genes in the ethanol-producing pathway. Ethanol was added in continuous fermentation to provide an ethanol concentration of either 17 or 27 g/L, triggering sustained oscillations in the cell growth rate. Cell concentrations oscillated in-phase with ethanol and acetate concentrations. The amplitude of oscillations depended on the concentration of ethanol in the fermentor. Through multiple oscillatory cycles, the yield (Y(P/S)) and concentration of ethanol decreased, while production of acetate increased. These results suggest that KO11 favorably adapted to improve growth by synthesizing more ATP though acetate production, and recycling NADH by producing more lactate and less ethanol. Implications of these results for strategies to improve ethanol production are described.


Gastrointestinal malformations in two infants born to women with hyperthyroidism untreated in the first trimester.
We report two infants with gastrointestinal anomalies: one with esophageal atresia and tracheo-esophageal fistula and the other with biliary tree atresia, born to hyperthyroid women diagnosed and treated with methimazole after 14 weeks' gestation. Euthyroidism was documented in both infants. These cases raise the issue of whether untreated hyperthyroidism and not methimazole intake is the teratogen.


Summertime Tintinnid Community in the Surface Waters Across the North Pacific Transition Zone.
Located from 35° to 45° latitude in both hemispheres, the transition zone is an important region with respect to the planktonic biogeography of the sea. However, to the best of our knowledge, there have been no reports on the existence of a tintinnid community in the transition zone. In this research, tintinnids along two transects across the North Pacific Transition Zone (NPTZ) were investigated in summer 2016 and 2019. Eighty-three oceanic tintinnid species were identified, 41 of which were defined as common oceanic species. The common oceanic species were further divided into five groups: boreal, warm water type I, warm water type II, transition zone, and cosmopolitan species. Undella californiensis and Undella clevei were transition zone species. Other species, such as Amphorides minor, Dadayiella ganymedes, Dictyocysta mitra, Eutintinnus pacificus, Eutintinnus tubulosus, Protorhabdonella simplex, and Steenstrupiella steenstrupii, were the most abundant in the NPTZ but spread over a much larger distribution region. Species richness showed no obvious increase in the NPTZ. Boreal, transition zone, and warm water communities were divided along the two transects. Tintinnid transition zone community mainly distributed in regions with water temperatures between 15 and 20°C. The tintinnid lorica oral diameter size classes were dominated by the 24-28 μm size class in three communities, but the dominance decreased from 66.26% in the boreal community to 48.85% in the transition zone community and then to 22.72% in the warm water community. Our research confirmed the existence of tintinnid transition zone species and community. The abrupt disappearance of warm water type I species below 15°C suggested that this group could be used as an indicator of the northern boundary of the NPTZ.


Cooperative hunting roles among taï chimpanzees.
All known chimpanzee populations have been observed to hunt small mammals for meat. Detailed observations have shown, however, that hunting strategies differ considerably between populations, with some merely collecting prey that happens to pass by while others hunt in coordinated groups to chase fast-moving prey. Of all known populations, Taï chimpanzees exhibit the highest level of cooperation when hunting. Some of the group hunting roles require elaborate coordination with other hunters as well as precise anticipation of the movements of the prey. The meat-sharing rules observed in this community guarantee the largest share of the meat to hunters who perform the most important roles leading to a capture. The learning time of such hunting roles is sometimes especially long. Taï chimpanzee males begin hunting monkeys at about age 10. The hunters' progress in learning the more sophisticated hunting roles is clearly correlated with age; only after 20 years of practice are they able to perform them reliably. This lengthy learning period has also been shown in some hunter-gatherer societies and confirms the special challenge that hunting represents.


Substrate-product analogue inhibitors of isoleucine 2-epimerase from Lactobacillus buchneri by rational design.
A rational approach that may be applied to a broad class of enzyme-catalyzed reactions to design enzyme inhibitors affords a powerful strategy, facilitating the development of drugs and/or molecular probes of enzyme mechanisms. A strategy for the development of substrate-product analogues (SPAs) as inhibitors of racemases and epimerases is elaborated using isoleucine 2-epimerase from Lactobacillus buchneri (LbIleE) as a model enzyme. LbIleE catalyzes the PLP-dependent, reversible, racemization or epimerization of nonpolar amino acids at the C-2 position. The enzyme plays an important role in the biosynthesis of branched-chain d-amino acids and is a potential target for the development of antimicrobial agents. 3-Ethyl-3-methyl-l-norvaline (Ki = 2.9 ± 0.2 mM) and 3-ethyl-3-methyl-d-norvaline (Ki = 1.5 ± 0.2 mM) were designed as SPAs based on the movement of the sec-butyl side chain of the substrate l-Ile during catalysis, and were competitive inhibitors with binding affinities exceeding that of l-Ile by 1.3- and 2.5-fold, respectively. Surprisingly, these compounds were not substrates, but the corresponding compounds lacking the 3-methyl group were substrates. Unlike serine, glutamate, and proline racemases, which exhibit stringent steric requirements at their active sites, the active site of LbIleE was amenable to binding bulky SPAs. Moreover, LbIleE bound the SPA 2,2-di-n-butylglycine (Ki = 11.0 ± 0.2 mM) as a competitive inhibitor, indicating that the hydrophobic binding pocket at the active site was sufficiently plastic to tolerate gem-dialkyl substitution at the α-carbon of an amino acid. Overall, these results reveal that amino acid racemases/epimerases are amenable to inhibition by SPAs provided that they possess a capacious active site.


Comparison of five rep-PCR genomic fingerprinting methods for differentiation of fecal Escherichia coli from humans, poultry and wild birds.
The development of a methodology to identify the origin of fecal pollution is important both for assessing the degree of risk posed to public health and for developing strategies to mitigate the environmental loading of pathogens associated with waterborne disease transmission. Five rep-PCR genomic fingerprinting methods, such as rep-PCR, enterobacterial repetitive intergenic consensus (ERIC)-PCR, ERIC2-PCR, BOX-PCR and (GTG)(5)-PCR, were assessed for their potential in differentiation of 232 fecal Escherichia coli isolates obtained from humans, poultry (chicken, duck and turkey) and wild birds (Canada goose and gull). Based on the results of cluster analysis and discriminant function analysis, (GTG)(5)-PCR was found to be the most suitable method for molecular typing of fecal E. coli, followed by BOX-PCR, REP-PCR, ERIC-PCR and ERIC2-PCR. A discriminant function analysis of (GTG)(5)-PCR fingerprints showed that 94.1%, 79.8%, 80.5%, 74.4%, 86.7% and 88.6% of turkey, chicken, duck, Canada goose, gull and human E. coli isolates were classified into the correct host group, respectively. Subsequently, (GTG)(5)-PCR was tested for its ability to track the origin of 113 environmental E. coli isolated from natural pond water. In conclusion, the (GTG)(5)-PCR genomic fingerprinting method can be considered as a promising genotypic tool for epidemiological surveillance of fecal pollution in aquatic environments.


Pathologic features of horses given avirulent equine arteritis virus intramuscularly.
Twenty horses that were seronegative for equine arteritis virus antibodies were inoculated IM with live equine arteritis virus vaccine. The inoculation did not cause clinical signs of disease. A mild, transient febrile reaction developed in 6 horses, 3 of which were in poor condition before inoculation. Six horses, 2 of which were in poor condition before inoculation, experienced mild lymphopenia. Necropsy revealed mild lesions in the lymph nodes of 6 horses (3 of which were in poor condition before inoculation). Maximum concentrations of virus were detected in the lymph nodes and were consistently present from postvaccination day 3 through 8. Lesser concentrations of virus were detected in the spleen of 5 horses, liver and kidney of 4, abdominal fluid of 3, pleural fluid of 2, and lungs and urine of 1, between postvaccination days 3 and 7. Virus was not detected in the brain, nasal tract, or serum of any of the horses.


Sintilimab plus fluorouracil, leucovorin, oxaliplatin and docetaxel regimen as neoadjuvant therapy for resectable gastric cancer and biomarker exploration.
At present, preoperative chemotherapy is the standard of care for the neoadjuvant treatment of potentially resectable gastric cancer (GC). However, because the efficacy and prognosis are not ideal, curative effects for this population are unsatisfactory. With the development of immune checkpoint inhibitors, the results of a few encouraging early trials of immunotherapeutic agents as neoadjuvant therapies for resectable GC have been reported. However, markers of the efficacy of immune checkpoint inhibitors remain unclear. This prospective single-center, single-arm observational study was designed to evaluate the efficacy of sintilimab plus the fluorouracil, leucovorin, oxaliplatin and docetaxel regimen as a neoadjuvant treatment for localized GC. More importantly, this work assesses multiple dimensions and include ctDNA, the immune microenvironment and intestinal microbiome to explore correlations between biomarkers and neoadjuvant therapeutic efficacy. Clinical trial registration: ChiCTR2200061629 (www.chictr.org.cn/index.aspx).


The Demand-Control Model as a Predictor of Depressive Symptoms-Interaction and Differential Subscale Effects: Prospective Analyses of 2212 German Employees.
Testing assumptions of the widely used demand-control (DC) model in occupational psychosocial epidemiology, we investigated (a) interaction, i.e., whether the combined effect of low job control and high psychological demands on depressive symptoms was stronger than the sum of their single effects (i.e., superadditivity) and (b) whether subscales of psychological demands and job control had similar associations with depressive symptoms. Logistic longitudinal regression analyses of the 5-year cohort of the German Study of Mental Health at Work (S-MGA) 2011/12-2017 of 2212 employees were conducted. The observed combined effect of low job control and high psychological demands on depressive symptoms did not indicate interaction (RERI = -0.26, 95% CI = -0.91; 0.40). When dichotomizing subscales at the median, differential effects of subscales were not found. When dividing subscales into categories based on value ranges, differential effects for job control subscales (namely, decision authority and skill discretion) were found (p = 0.04). This study does not support all assumptions of the DC model: (1) it corroborates previous studies not finding an interaction of psychological demands and job control; and (2) signs of differential subscale effects were found regarding job control. Too few prospective studies have been carried out regarding differential subscale effects.


Lymphocyte 5'-nucleotidase activities in normal dogs and in dogs with malignant lymphoma.
Plasma membrane 5'-nucleotidase (5'NT) activity was assayed in lymph node lymphocytes from seven normal control dogs and in malignant lymphocytes from 25 dogs with lymphoma. The lymphoid tumors were classified according to the NCI Working Formulation into five histologic subtypes. When compared with normal controls significantly lower 5'NT activities were found in the lymphoblastic, small lymphocytic and diffuse large (noncleaved) subtypes while no significant differences were observed in the immunoblastic or small noncleaved groups. In addition, dogs with hypercalcemia or paraproteinemia had decreased 5'NT levels. However, no significant differences were found between prognostic groups or histologic subtypes of the NCI classification. In conclusion, canine lymphoma subtypes had generally decreased 5'NT activities which appeared to reflect the B or T cell lineage, degree of maturation and enzyme activity of the cell of origin.


Wheeled mobility: factors influencing mobility and assistive technology in veterans and servicemembers with major traumatic limb loss from Vietnam war and OIF/OEF conflicts.
Returning wounded veterans and servicemembers to their highest level of function following traumatic injury is a priority of the Departments of Defense and Veterans Affairs. We surveyed 245 veterans from the Vietnam war and 226 servicemembers and veterans from Operation Iraqi Freedom/Operation Enduring Freedom (OIF/OEF) conflicts with at least one major traumatic lower-limb loss to determine their use of mobility assistive technology (AT) and patterns of limb abandonment. Prosthetic device use without wheelchair use is found in 50.5% of Vietnam and 42.8% of OIF/OEF groups. Prostheses and supplementary wheelchairs are used by Vietnam (32%) and OIF/OEF (53%) groups (p < 0.01). Exclusive wheelchair use is more frequent in the Vietnam group (18%) than in the OIF/OEF group (4.0%, p < 0.01). In Vietnam participants, multivariate analysis found that multiple-limb loss (adjusted odds ratio [AOR] = 14.5; 95% confidence interval [CI] 5.5-38.5), bilateral lower-limb loss (AOR = 12.7; 95% CI 6.2-26.1), and number of comorbidities (AOR = 1.3; 95% CI 1.2-1.5) are associated with increased likelihood of wheelchair use. In OIF/OEF participants, bilateral lower-limb loss (AOR = 29.8; 95% CI 11.0-80.7), multiple-limb loss (AOR = 16.3; 95% CI 3.1-85.3), cumulative trauma disorder (AOR = 2.4; 95% CI 1.2-4.9), and number of combat injuries (AOR = 1.4; 95% CI 1.2-1.7) are associated with wheelchair use. Combined use of different types of mobility ATs promotes improved rehabilitation and ability to function.


Trimeric G proteins modulate the dynamic interaction of PKAII with the Golgi complex.
The Golgi complex represents a major subcellular location of protein kinase A (PKA) concentration in mammalian cells where it has been previously shown to be involved in vesicle-mediated protein transport processes. We have studied the factors that influence the interaction of PKA typeII subunits with the Golgi complex. In addition to the cytosol, both the catalytic (Calpha) and regulatory (RIIalpha) subunits of PKAII were detected at both sides of the Golgi stack, particularly in elements of the cis- and trans-Golgi networks. PKAII subunits, in contrast, were practically absent from the middle Golgi cisternae. Cell treatment with either brefeldin A, AlF(4-) or at low temperature induced PKAII dissociation from the Golgi complex and redistribution to the cytosol. This suggested the existence of a cycle of association/dissociation of PKAII holoenzyme to the Golgi. The interaction of purified RIIalpha with Golgi membranes was studied in vitro and found not to be affected by brefeldin A while it was sensitive to modulators of heterotrimeric G proteins such as AlF(4-), GTPgammaS, beta(gamma) subunits and mastoparan. RII(alphaa) binding was stimulated by recombinant, myristoylated Galpha(i3) subunit and inhibited by cAMP. Pretreatment of Golgi membranes with bacterial toxins known to catalyze ADP-ribosylation of selected Galpha subunits also modified RIIalpha binding. Taken together the data support a regulatory role for Golgi-associated Galpha proteins in PKAII recruitment from the cytosol.


Intravitreal injection of cephaloridine in the treatment of endophthalmitis.
Ocular toxic effects of intravitreal injection of cephaloridine in rabbits was evaluated, and none were produced in doses of 0.25 mg or less. Clinically, intravitreal injection of cephaloridine in doses of 5 mg or more produced small punctate hemorrhages in the vascularized portions of the rabbit retina within two days. With doses of 2.5 mg or less, no clinical changes were seen during the two weeks of observation. Histologically, one day following intravitreal injection of 0.5 to 10 mg of cephaloridine there was clumping of the outer segments of the photoreceptors and destruction of the retinal pigment epithelium. Ten milligrams of cephaloridine intravitreally injected caused definite electroretinogram (ERG) changes ten minutes after injection. After 24 hours, the ERG was extinguished. An experimentally induced Escherichia coli endophthalmitis was successfully treated after six hours by intravitreal injection of cephaloridine.


Splenic and liver trauma in children.
These data indicate that most children with blunt injury to the liver and spleen can be treated conservatively. The fact that nonoperative management is a successful method of therapy limits the usefulness of DPL in most stable pediatric patients with abdominal trauma. Recovery of blood by DPL will neither influence the decision to operate nor yield information concerning which organ is injured. The abdominal CT scan is the most sensitive diagnostic method. In patients with associated head trauma, the head CT and abdominal CT both can be obtained during the same visit to the radiology suite. DPL may be useful in the unstable patient to be sure the abdomen is the site of bleeding before starting an emergency laparotomy and occasionally, in more stable patients with ongoing abdominal pain, to rule out an associated bowel injury with perforation (e.g., recovery of bilious or feculant material). Insight into the contemporary management of splenic and liver injuries in children is important. In the 1980s, a laparotomy safely can be avoided in many cases with reduction of patient morbidity, length of hospital stay and cost of hospitalization.


Electrically-Driven Soft Fluidic Actuators Combining Stretchable Pumps With Thin McKibben Muscles.
Soft wearable robots could provide support for lower and upper limbs, increase weight lifting ability, decrease energy required for walking and running, and even provide haptic feedback. However, to date most of wearable robots are based on electromagnetic motors or fluidic actuators, the former being rigid and bulky, the latter requiring external pumps or compressors, greatly limiting integration and portability. Here we describe a new class of electrically-driven soft fluidic muscles combining thin, fiber-like McKibben actuators with fully Stretchable Pumps. These pumps rely on ElectroHydroDynamics, a solid-state pumping mechanism that directly accelerates liquid molecules by means of an electric field. Requiring no moving parts, these pumps are silent and can be bent and stretched while operating. Each electrically-driven fluidic muscle consists of one Stretchable Pump and one thin McKibben actuator, resulting in a slender soft device weighing 2 g. We characterized the response of these devices, obtaining a blocked force of 0.84 N and a maximum stroke of 4 mm. Future work will focus on decreasing the response time and increasing the energy efficiency. Modular and straightforward to integrate in textiles, these electrically-driven fluidic muscles will enable soft smart clothing with multi-functional capabilities for human assistance and augmentation.


The MiniArc sling system in the treatment of female stress urinary incontinence.
To assess the technical feasibility of a new mini-invasive sling procedure (MiniArc) and present short-term results in the treatment of female urinary incontinence. A total of 97 women with mixed or stress urinary incontinence (SUI) were treated by placement of the new single-incision sling. Pelvic organ prolapse was graded using the POP-Q system (pelvic organ prolapse quantification system). Preoperative workup included urodynamic evaluation, cough stress test and introital ultrasound. Postoperatively, introital ultrasound was performed to determine residual urine and check tape position. Quality of life was measured using King's Health Questionnaire. A voiding diary and pad count served to verify the patients' subjective complaints. The MiniArc single-incision sling procedure was the initial intervention in 37 (38.2%) patients and the second intervention in 60 (61.7%) patients with recurrent incontinence. The cough stress test was negative in 79 (83.1%) women 6 weeks after the sling procedure and in 74 (77.8%) at 12 months. De novo urge occurred in 32 (36.8%) women. Quality of life was significantly improved at 12-month follow-up in 65 (69.1%) patients (p < 0.001). The number of pads decreased significantly from 2.2 to 0.6 (p < 0.001) after the procedure. One patient developed an hematoma and bladder perforation occurred in another. Our short-term clinical results suggest that the MiniArc is a safe and effective minimally invasive sling procedure for treating female SUI. Randomized comparative controlled trials and long-term results are still required to define the role of the new sling system in comparison to established mid-urethral tape techniques for treating incontinence.


Multi-wavelength source using low drive-voltage amplitude modulators for optical communications.
A simple and cost-effective technique for generating a flat, square-shaped multi-wavelength optical comb with 42.6 GHz line spacing and over 0.5 THz of total bandwidth is presented. A detailed theoretical analysis is presented, showing that using two concatenated modulators driven with voltages of 3.5 V(pi) are necessary to generate 11 comb lines with a flatness below 2dB. This performance is experimentally demonstrated using two cascaded Versawave 40 Gbit/s low drive voltage electro-optic polarisation modulators, where an 11 channel optical comb with a flatness of 1.9 dB and a side-mode-suppression ratio (SMSR) of 12.6 dB was obtained.


Prognostic significance of p53 immunoexpression in the survival of oral squamous cell carcinoma patients treated with surgery and neoadjuvant chemotherapy.
p53 status is a key biomarker for a variety of cancer types. However, it remains controversial whether p53 is an effective biomarker in oral squamous cell carcinoma (OSCC), particularly with regard to its prognostic value for OSCC patients with combinational treatment. The aim of the current study was to evaluate the prognostic potential of p53 immunoexpression in samples from OSCC patients treated with surgery only or surgery and neoadjuvant chemotherapy. p53 expression was assessed immunohistochemically in biopsy tissues from 44 OSCC patients with a mean follow-up of 35.6 months. Correlations between p53 status, tumor size (T-classification), lymph node status (N-classification) and clinical outcome were analyzed. It was observed that p53-positive and N0 cases correlated with higher 5-year survival rates in cases treated with surgery alone (P=0.017 and P=0.03, respectively), while in cases with neoadjuvant chemotherapy, p53 status and lymph node status did not exhibit prognostic significance. Tumor size showed no prognostic value in cases receiving surgery alone or in those with neoadjuvant chemotherapy. The present results demonstrated for the first time that p53 immunohistochemical expression correlates with a good prognosis in OSCC patients receiving surgery alone. In conclusion, p53 immunohistochemical expression and lymph node status may serve as prognostic markers for the survival of OSCC patients receiving surgery only, but not for patients undergoing surgery and neoadjuvant chemotherapy treatment.


Properties of antigenomic hepatitis delta virus ribozyme cis- and trans- analogs.
A series of permuted variants of antigenomic HDV ribozyme and trans-acting variants were constructed. The catalytic activity study of the ribozymes has shown that all the variants were capable of self-cleaving with equally biphasic kinetics. Ribonuclease and Fe(II)-EDTA cleavage have provided evidence that all designed ribozymes fold according to the pseudoknot model and the conformations of the initial and cleaved ribozyme are different. A scheme of HDV ribozyme self-cleavage reaction was suggested. The role of hydrogen bonds in the reaction was evaluated by substitution of ribose in the ribozyme for deoxyribose. It was found that the 2'-OH group of U23 and C27 is critical for the reaction to occur; the 2'-OH group of U32 and U39 is important, while 2'-OH groups of other nucleotides of loop 3, stem 4 and stem 1 are unimportant for the cleavage activity.


Streptococcus gordonii modulates Candida albicans biofilm formation through intergeneric communication.
The fungus Candida albicans colonizes human oral cavity surfaces in conjunction with a complex microflora. C. albicans SC5314 formed biofilms on saliva-coated surfaces that in early stages of development consisted of approximately 30% hyphal forms. In mixed biofilms with the oral bacterium Streptococcus gordonii DL1, hyphal development by C. albicans was enhanced so that biofilms consisted of approximately 60% hyphal forms. Cell-cell contact between S. gordonii and C. albicans involved Streptococcus cell wall-anchored proteins SspA and SspB (antigen I/II family polypeptides). Repression of C. albicans hyphal filament and biofilm production by the quorum-sensing molecule farnesol was relieved by S. gordonii. The ability of a luxS mutant of S. gordonii deficient in production of autoinducer 2 to induce C. albicans hyphal formation was reduced, and this mutant suppressed farnesol inhibition of hyphal formation less effectively. Coincubation of the two microbial species led to activation of C. albicans mitogen-activated protein kinase Cek1p, inhibition of Mkc1p activation by H(2)O(2), and enhanced activation of Hog1p by farnesol, which were direct effects of streptococci on morphogenetic signaling. These results suggest that interactions between C. albicans and S. gordonii involve physical (adherence) and chemical (diffusible) signals that influence the development of biofilm communities. Thus, bacteria may play a significant role in modulating Candida carriage and infection processes in the oral cavity.


COE1, an LRR-RLK responsible for commissural vein pattern formation in rice.
Leaf veins have a complex network pattern. Formation of this vein pattern has been widely studied as a model of tissue pattern formation in plants. To understand the molecular mechanism governing the vascular patterning process, we isolated the rice mutant, commissural vein excessive1 (coe1). The coe1 mutants had short commissural vein (CV) intervals and produced clustered CVs. Application of 1-N-naphthylphthalamic acid and brefeldin A decreased CV intervals, and application of 1-naphthaleneacetic acid increased CV intervals in wild-type rice; however, coe1 mutants were insensitive to these chemicals. COE1 encodes a leucine-rich repeat receptor-like kinase, whose amino acid sequence is similar to that of brassinosteroid-insensitive 1-associated receptor kinase 1 (BAK1), and which is localized at the plasma membrane. Because of the sequence similarity of COE1 to BAK1, we also examined the involvement of brassinosteroids in CV formation. Brassinolide, an active brassinosteroid, decreased the CV intervals of wild-type rice, and brassinazole, an inhibitor of brassinosteroid biosynthesis, increased the CV intervals of wild-type rice, but coe1 mutants showed insensitivity to these chemicals. These results suggest that auxin and brassinosteroids regulate CV intervals in opposite directions, and COE1 may regulate CV intervals downstream of auxin and brassinosteroid signals.


Distribution of hypertension and renal disease in Oregon.
Expecting to find agreement between the geographic distribution of hypertension and renal disease, we developed regional mortality rates for 1950-72 and prevalence rates for a Selective Service cohort born in 1939-41 and examined during 1957-69. For this purpose the State's counties were grouped into eight geographically homogeneous regions. The general decline in hypertension mortality was most pronounced in Portland, Oregon's major urban center. However, the decline halted during 1968-72 in the southern Cascade region which has become an area of relatively higher risk within the State. During these 23 years nephritis mortality fell, kidney infection mortality was stable, and both syndromes showed peak mortality in other, different regions of the State. The geographic pattern of hypertension prevalence among the draftee cohort resembled the 1963-67 hypertension mortality pattern, but more recent morbidity data are needed to confirm the southern Cascade region's recent change to a high-risk area. Of 529 draftees with diagnosed hypertension, only 35 percent of the cases were previously known, only 7 percent has had any previous treatment, and only 7 percent were associated with known renal conditions. Among 521 registrants with a history of renal disorders, the prevalence of hypertension was increased for all categories of renal disease but was significantly high only for those with a history of glomerulonephritis. To date in Oregon we have found no evidence that renal disorders are major determinants of hypertension morbidity or mortality.


Reporting of vertebral fractures on spine X-rays.
Vertebral fractures are the hallmark of osteoporosis, responsible for increased morbidity and mortality in post-menopausal women. However, two-thirds of vertebral fractures do not come to clinical attention. The aim of this study was to compare the identification of vertebral fractures on spine X-rays among rheumatologists. Study subjects were women aged 60-80 years having potential signs of vertebral fracture and visiting a rheumatologist. X-rays were performed according to standardized procedures. In 629 patients (among 824 included) at least one vertebral fracture was diagnosed, and the X-rays were then sent to a central facility where a semi-quantitative assessment of vertebral fracture was performed by a single rheumatologist trained for this evaluation. According to the vertebral level, kappa scores were between 0.20 to 0.77, i.e., below 0.6 from T4 to T7, and between 0.6 and 0.77 from T8 to L4. The false-negative fractures rate was 25.8% (and 15.7% of them were related to a numbering discrepancy). The rate of false positive fractures was 6.3%. At the patient level 6.8% had actually no fracture. This study shows that 25% of overall vertebral fractures are not diagnosed among patients considered as having at least one fracture. As a consequence, patients who require treatment to reduce fracture risk are not being properly identified.


Rheological and Curing Behavior of Acrylate-Based Suspensions for the DLP 3D Printing of Complex Zirconia Parts.
The present study demonstrates the possibility of fabricating zirconia parts with a complex shape and internal architecture using a low-cost stereolithography-based technique. One of the critical steps in ceramics stereolithography is the preparation of a photo-curable slurry with properties that fulfill specific requirements, such as having a low viscosity, high solids loading and appropriate curing characteristics. Slurries with different acrylic monomers and ceramic fillers were studied concerning their rheological and curing behavior. New formulations based on mono- and tri-functional acrylic monomers revealed the following excellent rheological properties: The viscosity of the mono-/tri-acrylate-based slurry with 75 wt.% of zirconia was 1.6 Pa·s at 30 s-1. Zirconia stabilized with 3 mol.% yttria was found to be more favorable than zirconia with 8 mol.% yttria for slurry preparation, because of its lower surface area and higher tapped density. It was shown that the cure depth of the suspensions was suitable for printing objects with a 50 µm layer thickness, good interlayers connection and surface finishing.


Endonasal transsphenoidal removal of tuberculum sellae meningiomas: technical note.
Tuberculum sellae meningiomas traditionally have been removed through a transcranial approach. More recently, the sublabial transsphenoidal approach has been used to remove such tumors. Here, we describe use of the direct endonasal transsphenoidal approach for removal of suprasellar meningiomas. Three women, aged 32, 34, and 55 years, each sought treatment for visual loss and headaches. In each patient, magnetic resonance imaging (MRI) showed a suprasellar mass causing optic chiasmal and optic nerve compression (average size, 2 x 2 cm). All three patients underwent tumor removal via an endonasal approach with the operating microscope. Suprasellar exposure was facilitated by removal of the posterior planum sphenoidale. Ultrasound was used to help define tumor location before dural opening. The extent of tumor removal was verified with angled endoscopes in all patients, and with intraoperative MRI in one patient. The surgical dural and bony defects were repaired in all patients with abdominal fat, titanium mesh, and 2 to 3 days of cerebrospinal fluid lumbar drainage. Nasal packing was not used. There were no postoperative cerebrospinal fluid leaks or meningitis. One patient required a reoperation 2 weeks after surgery to reduce the size of her fat graft, which was causing optic nerve compression; within 24 hours, her vision rapidly improved. At 3 months after surgery, all three patients had normal vision, no new endocrinopathy, and no residual tumor on MRI. At 10 months after surgery, one patient had a small asymptomatic tumor regrowth seen on MRI. The endonasal approach with the operating microscope appears to be an effective minimally invasive method for removing relatively small midline tuberculum sellae meningiomas. Intraoperative ultrasound, the micro-Doppler probe, and angled endoscopes are useful adjuncts for safely and completely removing such tumors. Longer follow-up is needed to monitor for tumor recurrence in these patients.


Clinical Characteristics, Treatment, and Outcomes of Critically Ill Patients With COVID-19: A Scoping Review.
A growing number of studies on coronavirus disease 2019 (COVID-19) are becoming available, but a synthesis of available data focusing on the critically ill population has not been conducted. We performed a scoping review to synthesize clinical characteristics, treatment, and clinical outcomes among critically ill patients with COVID-19. Between January 1, 2020, and May 15, 2020, we identified high-quality clinical studies describing critically ill patients with a sample size of greater than 20 patients by performing daily searches of the World Health Organization and LitCovid databases on COVID-19. Two reviewers independently reviewed all abstracts (2785 unique articles), full text (218 articles), and abstracted data (92 studies). The 92 studies included 61 from Asia, 16 from Europe, 10 from North and South America, and 5 multinational studies. Notable similarities among critically ill populations across all regions included a higher proportion of older males infected and with severe illness, high frequency of comorbidities (hypertension, diabetes, and cardiovascular disease), abnormal chest imaging findings, and death secondary to respiratory failure. Differences in regions included newly identified complications (eg, pulmonary embolism) and epidemiological risk factors (eg, obesity), less chest computed tomography performed, and increased use of invasive mechanical ventilation (70% to 100% vs 15% to 47% of intensive care unit patients) in Europe and the United States compared with Asia. Future research directions should include proof-of-mechanism studies to better understand organ injuries and large-scale collaborative clinical studies to evaluate the efficacy and safety of antivirals, antibiotics, interleukin 6 receptor blockers, and interferon. The current established predictive models require further verification in other regions outside China.


Comparisons of three different methods for defining sarcopenia: An aspect of cardiometabolic risk.
Appraisal of muscle mass is important when considering the serious consequences of sarcopenia in an aging society. However, the associations between sarcopenia and its clinical outcomes might vary according to the method applied in its diagnosis. We compared the relationships between cardiometabolic risk parameters and sarcopenia defined according to three different diagnostic methods using dual-energy X-ray absorptiometry (DXA) and computed tomography (CT). Appendicular skeletal muscle mass (ASM) adjusted by height squared and BMI (ASM/height2 and ASM/BMI) measured using DXA and thigh muscle cross-sectional area (tmCSA) adjusted by weight (tmCSA/weight) measured using CT were used as indices of muscle mass. Sarcopenia was defined as two standard deviations below either the mean ASM/height2, ASM/BMI, or tmCSA/weight of a young reference group. ASM/BMI and tmCSA/weight showed a negative relationship with several components of metabolic syndrome and HOMA-IR, whereas ASM/height2 was positively associated with theses cardiometabolic risk factors. Logistic regression analyses demonstrated that ASM/BMI-defined sarcopenia was significantly associated with increased HOMA-IR (P = 0.01) and prevalence of visceral obesity (P = 0.03) and metabolic syndrome (P = 0.025), while ASM/height2- and tmCSA/weight-defined sarcopenia were not. ASM/BMI-defined sarcopenia exhibits a closer relationship with cardiometabolic risk factors than does ASM/height2- or tmCSA/weight-defined sarcopenia.


Skin disease affecting the conservation of the western swamp tortoise (Pseudemydura umbrina)
To review the present position of the western swamp tortoise (Pseudemydura umbrina) as an endangered species and significant health issues affecting efforts to save it from extinction. A retrospective analysis of the husbandry, hospital and pathology records of the western swamp tortoise captive breeding program at Perth Zoo. In 1987 a captive breeding project was developed to prevent the extinction of the western swamp tortoise but an outbreak of a necrotising dermatitis in 1989 threatened the survival of the captive bred hatchlings. Less severe outbreaks occurred in 1990 and 1993, with isolated cases in between. Of 283 tortoises that were born in captivity or came into captivity from the wild, 37 (13.1%) were affected, comprising 37% of all males, 26% of all females and 13% of animals of unknown gender. Of the affected animals, 70% were less than 2 years of age and 29% were older. Males were 1.6 times more likely to be infected than females but this difference was not statistically significant (P = 0.27). Culture of the lesions consistently yielded unidentified Pseudomonas sp. Improved husbandry, such as strict maintenance of water quality and temperature conditions similar to that of the animal's natural habitat, and monitoring the health of individual tortoises have successfully controlled skin disease in the captive breeding of the western swamp tortoise.


Human immunodeficiency virus disease-related neutropenia and the risk of hospitalization for bacterial infection.
Neutropenia is common in patients with human immunodeficiency virus (HIV) disease. However, the degree of risk for serious bacterial infections associated with various levels of neutropenia in patients with HIV disease is not well defined. A retrospective analysis of databases containing demographic information for patients attending the San Francisco General Hospital HIV outpatient clinic, test results reported by the hospital's clinical laboratory, and the San Francisco General Hospital inpatient International Classification of Diseases, Ninth Revision (ICD-9) hospital discharge diagnosis codes from October 1, 1992, through November 30, 1993. Risk window time periods were defined, encompassing dates that consecutive absolute neutrophil counts (ANCs) occurred in a single ANC stratum. One risk window at the lowest ANC stratum for each patient was analyzed for hospitalizations with ICD-9 codes indicating bacterial infections. A 5% random sample of medical records was reviewed for end point validation. Codes from ICD-9 had 98% and 96% positive and negative predictive values, respectively, for meeting National Institute of Allergy and Infectious Diseases Division of AIDS [acquired immunodeficiency syndrome] clinical trial end point definitions for bacterial infections. Among 2047 evaluable patients, a significant increase in the incidence of hospitalization for serious bacterial infections was observed for those in the ANC strata of 500 to 749 X 10(6)/L and below. The 95% confidence intervals for the incidence of hospitalization associated with each ANC stratum below 500 X 10(6)/L did not overlap with that for any stratum of 750 X 10(6)/L or higher (22-117 vs 0.4-19 patient hospitalizations per 10000 days at risk, respectively). A multivariate analysis revealed only the severity and duration of neutropenia and black race to be significant end point predictors. Among 2047 patients with HIV disease, significantly higher risks of hospitalization for bacterial infections were associated with ANCs lower than 750 X 10(6)/L, especially for ANCs lower than 500 X 10(6)/L.


Increased sodium ion conductance through nicotinic acetylcholine receptor channels in PC12 cells exposed to nerve growth factors.
22Na+ uptake in response to cholinergic stimulation was measured in PC12 cells grown in media containing either beta-nerve growth factor (NGF) from mouse submaxillary gland or a component of Bungarus multicinctus venom (fraction 9B) which has nerve growth factor activity. Our results showed that each of these nerve growth factors increased the Na+ conductance capacity of ACh receptor-linked ion channels in PC12 cells. Compared to untreated cells, Na+ uptake was enhanced by about 40% in cells exposed for 24 to 30 hr to either 50 ng/ml of beta-NGF or to 5 micrograms/ml of fraction 9B. This cholinergically stimulated Na+ flux increased with time and reached a level 2- to 3-fold higher than that of untreated cells after 4 to 6 days' exposure to NGFs. The generation of neurites in response to NGFs from either source occurred with the same time course as the increase in Na+ channel conductance. Growth factor-enhanced Na+ uptake remained for several days after NGFs were removed from cells and neurites were lost. Na+ flux into untreated and into 9B- or beta-NGF-treated cells was inhibited to a greater extent by a nicotinic than by a muscarinic antagonist. Dibutyryl cyclic AMP did not cause an increase in Na+ uptake in response to cholinergic agonists at concentrations which stimulated neurite-like process formation, and dibutyryl cyclic AMP did not potentiate the effects of NGFs.


The impact of neuraminidase on apoptosis in cultures of blood lymphocytes isolated from rats bearing morris hepatoma.
Lymphocytes were obtained by heart-punction from rats bearing Morris hepatoma. In the short term, 18-hour cultures of these lymphocytes exhibited a significantly higher amount of apoptotic cells than lymphocyte cultures from the healthy, control animals. Neuraminidase, injected into the caudal vein of the rats with Morris hepatoma, caused a marked lowering in the amount of apoptotic blood-lymphocytes and an elevation of the amount of viable cells. The possible mechanism of neuraminidase preventing the apoptosis of blood-circulating lymphocytes in tumour hosts is discussed herein.


What is the evidence for platelet transfusion in perioperative settings?
Platelet transfusions are widely administered to restore perioperative haemostasis in haemorrhagic patients; however, the role of platelet transfusion is not well understood and administration is often based on empiric data. This review aims to explore consensus regarding platelet transfusion trigger, dose and how the haemostatic efficacy of platelet transfusion was assessed for the treatment of perioperative bleeding. A literature search was carried out using MEDLINE (PubMed) on 28 February 2017, to identify publications reporting the effect of platelet transfusion in relation to triggers, dose and assessment of haemostatic efficacy in bleeding patients in a perioperative setting. Eight publications were identified across a variety of settings, covering both prophylactic and therapeutic platelet transfusion in adult patients; the majority of the reports were in cardiac surgery. A high degree of variability was observed in the published studies, with only 50% of articles specifying a trigger for platelet transfusion. The most commonly used trigger was platelet count (25% of publications), with no consensus identified regarding the platelet count values used as triggers. Doses reported per transfusion varied from 1 to 12 units, and outcome measures were mixed, although the majority of publications (63%) assessed the requirement for transfusion with other blood products. The lack of consensus in published studies hinders our ability to draw conclusions regarding platelet transfusion and highlights the need for further studies to assess the appropriate dose and triggers for use in perioperative patients.


Intratumoral expression of a fusogenic membrane glycoprotein enhances the efficacy of replicating adenovirus therapy.
We describe here a novel strategy to enhance the in vivo efficacy of replicating adenovirus therapy, using coinjection of plasmid DNA encoding a fusogenic viral glycoprotein. The combination of fusogenic membrane glycoprotein (FMG)-induced tumor cell fusion and infection with replicating adenovirus effectively treats even large established tumors at doses of plasmid DNA and virus that alone are ineffective. Adenoviral infection appears to increase the transduction of the tumor cells to a modest degree thereby boosting the FMG-mediated component of the therapy. Simultaneously, syncytial formation enhances the therapeutic effects of viral infection by increasing spread of adenoviral particles through the tumor cell population and by increasing titer of virus released from the tumor cells. This effect is due probably to release of intracellular viral particles upon tumor cell death and also to increased levels of E1A protein within syncytia, whose increased metabolic rate is associated with enhanced levels of protein expression. Cotransduction of tumor cells with replicating adenovirus and FMG-expressing vectors could either be combined within single replicating vectors or could be used in strategies using separate administration of two components, both at lower doses than required for either therapy alone.


Cytokine signatures of transformed B cells with distinct Epstein-Barr virus latencies as a potential diagnostic tool for B cell lymphoma.
Immunocompromised individuals, including those infected with human immunodeficiency virus (HIV), are at increased risk of Epstein-Barr virus (EBV)-associated aggressive B cell malignancies such as Burkitt's lymphoma (BL) or diffuse large B cell lymphoma (DLBCL). Differential diagnosis of these lymphomas requires histopathological, immunohistochemical and cytogenetic assessments. Rapid, less invasive approaches to the diagnosis of EBV-associated B cell lymphomas are needed. Here, high-throughput cytokine profiling of BL cell lines and EBV-transformed B lymphoblastoid cell lines (B-LCL), representing DLBCL, was carried out. By monitoring the production of 42 different cytokines, unique cytokine signatures were identified for BL and B-LCL/DLBCL. The BL cells produced interleukin (IL)-10, 10 kDa interferon gamma-induced protein (IP-10)/CXCL10, macrophage-derived chemokine (MDC)/CCL22, macrophage inflammatory protein (MIP)-1α/CCL3 and MIP-1β/CCL4. In addition to these five cytokines, the cytokine signature of B-LCL/DLBCL cells included IL-8/CXCL8, IL-13, platelet-derived growth factor (PDGF)-AA, and regulated upon activation, normal T cell expressed and secreted (RANTES)/CCL5. Epstein-Barr virus latency was responsible for the increased production of IL-10, MDC/CCL22 and MIP-1α/CCL3 in BL cells, suggesting that EBV-mediated BL-genesis involves these three cytokines. These results suggest that high-throughput cytokine profiling might be a valuable tool for the differential diagnosis and might deepen our understanding of the pathogenesis of EBV-associated B cell malignancies.


The biochemical inhibition mode of bredinin-5'-monophosphate on DNA polymerase beta.
We reported previously [T. Horie, Y. Mizushina, M. Takemura, F. Sugawara, A. Matsukage, S. Yoshida, K. Sakaguchi, Int. J. Mol. Med., 1 (1998) 83-90.] that a 5'-monophosphate form (breMP) of bredinin, which has been used clinically as an immunosuppressive drug, selectively suppressed the activities of mammalian DNA polymerase alpha (pol. alpha) and beta (pol. beta). In a preliminary study of the action mode, for pol. beta, breMP acted by competing with, unexpectedly, not only the substrate but also with the template-primer. The mode might be attributable to the structure and function of pol. beta itself. We therefore investigated the biochemical inhibition mode of pol. beta in more detail by using two pol. beta fragments which were proteolytically separated into the template-primer-binding domain and the catalytic domain. BreMP inhibited only the catalytic activity of the catalytic domain fragment, and could not bind to the template-primer-binding domain fragment, suggesting that it directly competes with the substrate at its binding site of the catalytic domain, and indirectly, but simultaneously and competitively disturbs the template-primer incorporation into the template-primer-binding domain.


The P2X7 receptor and Pannexin-1 are both required for the promotion of multinucleated macrophages by the inflammatory cytokine GM-CSF.
The P2X(7) receptor (P2X(7)R), an ATP-gated ion channel, has been implicated in the process of cell-to-cell fusion into multinucleated macrophages (MA), but its contribution to MA fusion driven by physiological/pathological stimuli is not clearly established. Based on several lines of evidence, we demonstrate that P2X(7)R is critical for the induction of multinucleated MA by the inflammatory cytokine GM-CSF: 1) pharmacological inhibition of P2X(7)R with oxidized ATP (oATP), KN-62, and the selective antagonist A740003 abrogated GM-CSF action on rat alveolar MA and murine peritoneal MA; 2) a murine J774 P2X(7) low MA clone, selected for defective P2X(7)R function, was unresponsive; 3) MA from mice lacking P2X(7)R failed to respond to GM-CSF, in contrast to wild-type. GM-CSF also stimulated ATP-induced membrane permeabilization in J774 P2X(7) high MA and rat alveolar MA, an effect absent in the P2X(7) low MA clone and inhibited by the P2X(7) blockers oATP and KN-62. Notably, the stimulatory effects of GM-CSF on pore formation and MA fusion were both inhibited by blocking functional Pannexin-1 (Panx-1), and GM-CSF failed to stimulate MA fusion in cells from Panx-1 knockout mice. We provide further evidence that extracellular ATP release from peritoneal MA is dependent on P2X(7) but not on Panx-1 expression and that its metabolism to adenosine mediates P2X(7)-dependent MA fusion. These data demonstrate that both P2X(7) and Panx-1 are required for GM-CSF promotion of MA fusion but likely act independently through different signaling pathway(s).


Early postzygotic mutations contribute to de novo variation in a healthy monozygotic twin pair.
Human de novo single-nucleotide variation (SNV) rate is estimated to range between 0.82-1.70×10(-8) mutations per base per generation. However, contribution of early postzygotic mutations to the overall human de novo SNV rate is unknown. We performed deep whole-genome sequencing (more than 30-fold coverage per individual) of the whole-blood-derived DNA samples of a healthy monozygotic twin pair and their parents. We examined the genotypes of each individual simultaneously for each of the SNVs and discovered de novo SNVs regarding the timing of mutagenesis. Putative de novo SNVs were validated using Sanger-based capillary sequencing. We conservatively characterised 23 de novo SNVs shared by the twin pair, 8 de novo SNVs specific to twin I and 1 de novo SNV specific to twin II. Based on the number of de novo SNVs validated by Sanger sequencing and the number of callable bases of each twin, we calculated the overall de novo SNV rate of 1.31×10(-8) and 1.01×10(-8) for twin I and twin II, respectively. Of these, rates of the early postzygotic de novo SNVs were estimated to be 0.34×10(-8) for twin I and 0.04×10(-8) for twin II. Early postzygotic mutations constitute a substantial proportion of de novo mutations in humans. Therefore, genome mosaicism resulting from early mitotic events during embryogenesis is common and could substantially contribute to the development of diseases.


A novel 3-D cuprous iodide polymer with a high Cu/I ratio.
A novel 3-D cuprous iodide polymer [CuI(μ6-L)]n with a Cu/I ratio as high as 3 (1, H2L = 1,3-bis(2H-tetrazol-5-yl)benzene) has been hydrothermally synthesized. The 3-D framework of 1 is constructed by the linkages of rare 1-D cationic aggregates [CuI2+]n and anionic bi-tetrazolate bridges. 1 shows unusually thermochromic luminescent and photocatalytic properties, and its density functional theory calculation has also been studied.