The role of palladin in actin organization and cell motility.
Palladin is a widely expressed protein found in stress fibers, focal adhesions, growth cones, Z-discs, and other actin-based subcellular structures. It belongs to a small gene family that includes the Z-disc proteins myopalladin and myotilin, all of which share similar Ig-like domains. Recent advances have shown that palladin shares with myotilin the ability to bind directly to F-actin, and to crosslink actin filaments into bundles, in vitro. Studies in a variety of cultured cells suggest that the actin-organizing activity of palladin plays a central role in promoting cell motility. Correlative evidence also supports this hypothesis, as palladin levels are typically up-regulated in cells that are actively migrating: in developing vertebrate embryos, in cells along a wound edge, and in metastatic cancer cells. Recently, a mutation in the human palladin gene was implicated in an unusually penetrant form of inherited pancreatic cancer, which has stimulated new ideas about the role of palladin in invasive cancer.


Degradation of rice bran hemicellulose by Paenibacillus sp. strain HC1: gene cloning, characterization and function of beta-D-glucosidase as an enzyme involved in degradation.
A bacterium (strain HC1) capable of assimilating rice bran hemicellulose was isolated from a soil and identified as belonging to the genus Paenibacillus through taxonomical and 16S rDNA sequence analysis. Strain HC1 cells grown on rice bran hemicellulose as a sole carbon source inducibly produced extracellular xylanase and intracellular glycosidases such as beta-D-glucosidase and beta-D-arabinosidase. One of them, beta-D-glucosidase, was further analyzed. A genomic DNA library of the bacterium was constructed in Escherichia coli and gene coding for beta-D-glucosidase was cloned by screening for beta-D-glucoside-degrading phenotype in E. coli cells. Nucleotide sequence determination indicated that the gene for the enzyme contained an open reading frame consisting of 1,347 bp coding for a polypeptide with a molecular mass of 51.4 kDa. The polypeptide exhibits significant homology with other bacterial beta-D-glucosidases and belongs to glycoside hydrolase family 1. Beta-D-Glucosidase purified from E. coli cells was a monomeric enzyme with a molecular mass of 50 kDa most active at around pH 7.0 and 37 degrees C. Strain HC1 glycosidases responsible for degradation of rice bran hemicellulose are expected to be useful for structurally determining and molecularly modifying rice bran hemicellulose and its derivatives.


The contribution of a covalently bound cofactor to the folding and thermodynamic stability of an integral membrane protein.
The factors controlling the stability, folding, and dynamics of integral membrane proteins are not fully understood. The high stability of the membrane protein bacteriorhodopsin (bR), an archetypal member of the rhodopsin photoreceptor family, has been ascribed to its covalently bound retinal cofactor. We investigate here the role of this cofactor in the thermodynamic stability and folding kinetics of bR. Multiple spectroscopic probes were used to determine the kinetics and energetics of protein folding in mixed lipid/detergent micelles in the presence and absence of retinal. The presence of retinal increases extrapolated values for the overall unfolding free energy from 6.3 ± 0.4 kcal mol(-1) to 23.4 ± 1.5 kcal mol(-1) at zero denaturant, suggesting that the cofactor contributes 17.1 kcal mol(-1) towards the overall stability of bR. In addition, the cooperativity of equilibrium unfolding curves is markedly reduced in the absence of retinal with overall m-values decreasing from 31.0 ± 2.0 kcal mol(-1) to 10.9 ± 1.0 kcal mol(-1), indicating that the folded state of the apoprotein is less compact than the equivalent for the holoprotein. This change in the denaturant response means that the difference in the unfolding free energy at a denaturant concentration midway between the two unfolding curves is only ca 3-6 kcal mol(-1). Kinetic data show that the decrease in stability upon removal of retinal is associated with an increase in the apparent intrinsic rate constant of unfolding, k(u)(H2O), from ~1 × 10(-16) s(-1) to ~1 × 10(-4) s(-1) at 25 °C. This correlates with a decrease in the unfolding activation energy by 16.3 kcal mol(-1) in the apoprotein, extrapolated to zero SDS. These results suggest that changes in bR stability induced by retinal binding are mediated solely by changes in the activation barrier for unfolding. The results are consistent with a model in which bR is kinetically stabilized via a very slow rate of unfolding arising from protein-retinal interactions that increase the rigidity and compactness of the polypeptide chain.


Development of orthogonal task designs in fMRI studies of higher cognition: the NIMH experience.
This paper chronicles one researcher's journey at the National Institute of Mental Health, exploring ways to understand the neural systems responsible for the cognitive sub-processes of working memory tasks. Both the opportunities and the pitfalls with applying the idea of cognitive subtraction to neuroimaging data were well-known from studies using positron emission tomography. We took advantage of the improved temporal resolution of fMRI with a delayed-recognition task and identified the time-courses of the different stages of the task (encoding, memory delay, and recognition test) as predictor variables in a multiple regression analysis. Because these signals were temporally independent, individual components of tasks could be contrasted with one another, rather than entire tasks, reducing the problem of violations of pure insertion in cognitive subtraction. This approach enabled us to draw more detailed conclusions about the neural systems of higher cognition and the organization of prefrontal cortex than had been possible before fMRI. Further enhancements and innovations over the last 20 years by a multitude of researchers across the field have greatly expanded this knowledge, but this approach called "orthogonal task design" has remained a fundamental component of many of these modern studies.


Sildenafil and T-1032, phosphodiesterase type 5 inhibitors, showed a different vasorelaxant property in the isolated rat aorta.
The vasorelaxant effects of sildenafil and T-1032 [methyl-2-(4-aminophenyl)-1,2-dihydro-1-oxo-7-(2-pyridinylmethoxy)-4-(3,4,5-trimethoxyphenyl)-3-isoquinoline carboxylate sulfate], two phosphodiesterase type 5 inhibitors, were examined in the isolated rat aorta. Sildenafil and T-1032, both of which have almost the same potency and selectivity regarding phosphodiesterase type 5 inhibitory activity, produced a similar, moderate, relaxation at 10(-10) to 10(-7) M (sildenafil: 66.8 +/- 13.7%; T-1032: 77.9 +/- 10.8% at 10(-7) M). However, sildenafil, but not T-1032, produced further relaxation at the higher concentrations (sildenafil: 102.0 +/- 0.6%; T-1032: 81.0 +/- 7.2% at 10(-4) M, P < 0.05). Sildenafil also produced a more potent relaxation than did T-1032 at the high concentrations (10(-5) and 10(-4) M) in endothelium-denuded aortic rings and in the presence of N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor (3 x 10(-4) M). Moreover, the high concentrations of sildenafil, but not of T-1032, caused a rightward shift of the concentration-response curve for calcium chloride in K(+)-depolarized endothelium-denuded preparations. In the ligand binding assay for the L-type Ca(2+) channels, the affinities of sildenafil at 10(-5) M for binding sites of nitrendipine and (--)-desmethoxyverapamil [(--)- D888] (35.2 +/- 3.3% and 35.8 +/- 1.9%, respectively) were higher than those of T-1032 (11.8 +/- 4.0% and -13.1 +/- 1.3%, respectively, P < 0.05). Regarding cyclic nucleotide levels, both phosphodiesterase type 5 inhibitors increased cGMP levels at 10(-6) M. However, sildenafil, but not T-1032, further increased cGMP levels at the higher concentrations (sildenafil: 15.7 +/- 2.7 pmol/mg protein; T-1032: 5.6 +/- 0.6 pmol/mg protein at 10(-4) M, P < 0.05). These results suggested that high concentrations of sildenafil had additional vasorelaxant properties through mechanisms other than phosphodiesterase type 5 inhibition. Sildenafil-induced relaxation appears to be due to inhibition of the external Ca(2+)-dependent cascade for contraction and/or to an increase in cGMP levels. In contrast, T-1032 only showed a vasorelaxant property due to phosphodiesterase type 5 inhibition. In conclusion, T-1032 appears to be a specific phosphodiesterase type 5 inhibitor compared with sildenafil and a useful compound to examine the physiological function of phosphodiesterase type 5.


Multiple myeloma in a mink handler following exposure to Aleutian disease.
Aleutian disease of mink is a viral illness characterized by systemic plasmocytosis and hypergammaglobulinemia. Some affected mink develop a monoclonal gammaglobulin spike and Bence-Jones proteinuria. A case of multiple myeloma in a mink handler with a 15-year history of exposure to Aleutian mink is presented. Previously reported cases of possible Aleutian disease (AD) in man are discussed and the pathogenesis of AD reviewed. Aleutian disease virus (ADV) may produce asymptomatic infection in exposed individuals. Available data suggest symptomatic disease in humans is extremely rare.


Are rigidity and tremor two sides of the same coin in Parkinson's disease?
In this research, we evaluated the inter-relation of tremor and rigidity in Parkinson's disease. We included the agonist and antagonist skeletal muscle models as well as the peripheral (spinal and long-loop reflexes) and central (basal ganglia and cortex) mechanisms and present a complete model. All of our simulation is developed in SIMULINK, MATLAB 7. Our results showed that (1) increased supplementary motor area output increases the rigidity and (2) rigidity and the tremor are correlated. Therefore, from our model it can be suggested that the simultaneous treatment of these two abnormal motor behaviors could be more beneficial. Experimental studies are needed to validate this computational model.


Urinary glycosaminoglycan levels following induced cystitis in monkeys.
Urinary glycosaminoglycan (GAG) levels were measured by the Whiteman assay in five monkeys following induction of an E. coli urinary tract infection. Urinary GAG levels rose as the infection developed and returned to baseline levels as the infection resolved. Elevated urinary GAG levels may be a marker for the tissue injury incurred by such infections and may offer insight into their pathophysiology.


Isolation and characterization of two new low-molecular-weight protein proteinase inhibitors from the granule-rich fraction of equine neutrophilic granulocytes.
A new species of protein proteinase inhibitors was detected in the granule-rich fraction of equine neutrophilic granulocytes. Five isoinhibitors were identified with a narrow enzyme specificity towards two microbial proteinases, e.g., proteinase K and subtilisin. Two isoinhibitors were purified and partially characterized. They had an Mr of 11,300 and 7400, respectively, and were resistant to perchloric acid and heat treatment at 100 degrees C for 20 min. The inhibitors retained their activity over a broad range of pH (1-9 and 1-12, respectively). The possible biological function of this species of protein proteinase inhibitors as defensins (= endogenous antibiotics) is tentatively discussed.


Ossiculoplasty using incus interposition: hearing results and analysis of the middle ear risk index.
To determine the immediate hearing result and the long-term stability of sculpted incus interposition in ossiculoplasty and evaluate the utility of the middle ear risk index in predicting hearing outcome in these cases. One hundred thirty-seven surgical patients. Review of 137 patients who underwent ossiculoplasty using autologous or homologous sculpted incus interposition. Ossiculoplasty using autologous or homologous sculpted incus interposition. Retrospective chart review, using the guidelines delineated by the Committee on Hearing and Equilibrium of the Academy of Otolaryngology-Head and Neck Surgery for the evaluation of results for the treatment of conductive hearing loss. The mean preoperative air bone gap was 26.8 dB, and the mean postoperative gap was 18.6 dB. Twenty-seven percent of patients were closed to within 10 dB, and 66.4% were brought to within 20 dB of the postoperative bone conduction line. Average time to the last postoperative audiometric testing was 15.8 months, with a range of 2 to 62 months. A mean air bone gap change of -0.2 dB was noted. Four patients had more than a 10 dB deterioration in conductive hearing loss. There were no cases of graft extrusion. Each ear operated upon in our series was fully scored using the middle ear risk index, and an index total was calculated. No statistical associations could be demonstrated in any group between the postoperative air bone gap and the middle ear risk index subcategories or total. Sculpted autologous or homologous incus interposition provides hearing success comparable with current allograft prosthesis studies, has a very low extrusion rate, and remains stable over time. We were not able to demonstrate an association between the middle ear risk index and hearing results in this subset of patients.


Defining the cost of the Egyptian lymphatic filariasis elimination programme.
Lymphatic filariasis (LF) is targeted for global elimination. LF elimination programmes in different countries, including Egypt, are supported financially by national and international agencies. The national programme in Egypt is based on mass drug administration (MDA) of an annual dose of a combination of 2 drugs (DEC and albendazole) to all endemic villages. This study aimed primarily to estimate the Total and Government costs of two rounds of MDA conducted in Egypt in 2000 and 2001, the average cost per person treated, and the cost share of the different programme partners. The Total costs reflect the overall annual costs of the MDA programme, and we defined Government costs as those expenditures made by the Egyptian government to develop, implement and sustain the MDA programmes. We used a generic protocol developed in coordination with the Emory Lymphatic Filariasis Support Center. Our study was concerned with all costs to the government, donors and other implementing parties. Cost data were retrospectively gathered from local, regional and national Ministry of Health and Population records. The total estimates for each governorate were based on data from a representative district for the governorate; these were combined with national programme data for a national estimate. The overall Total and Government costs for treating approximately 1,795,553 individuals living in all endemic villages in the year 2000 were USD 3,181,000 and USD 2,412,000, respectively. In 2001, the number of persons treated increased (29%) and the Total costs were USD 3,109,000 while Government costs were USD 2,331,000. In 2000, the average Total and Government costs per treated subject were USD 1.77 and USD 1.34, respectively, however, these costs decreased to USD 1.34 and USD 1.00, respectively in 2001. The coverage rate was 86.0% in 2000 and it increased to 88.0% in 2001. The Egyptian government provided 75.8% of all resources, as reflected in the Total cost estimates, and international agencies contributed the rest. Such data highlight both the commitment of the Egyptian government and the significance of the contributions of international bodies toward the LF elimination programme.


Caffeine prevents prostaglandin E1-induced disturbances in respiratory control in neonatal rats: implications for infants with critical congenital heart disease.
Continuous infusion of prostaglandin E1 (PGE1) is used to maintain ductus arteriosus patency in infants with critical congenital heart disease, but it can also cause central apnea suggesting an effect on respiratory neural control. In this study, we investigated whether 1) PGE1 inhibits the various phases of the acute hypoxic ventilatory response (HVR; an index of respiratory control dysfunction) and increases apnea incidence in neonatal rats; and 2) whether these changes would be reversible with caffeine pretreatment. Whole body plethysmography was used to assess the HVR and apnea incidence in neonatal rats 2 h following a single bolus intraperitoneal injection of PGE1 with and without prior caffeine treatment. Untreated rats exhibited a biphasic HVR characterized by an initial increase in minute ventilation followed by a ventilatory decline of the late phase (~5th minute) of the HVR. PGE1 had a dose-dependent effect on the HVR. Contrary to our hypothesis, the lowest dose (1 µg/kg) of PGE1 prevented the ventilatory decline of the late phase of the HVR. However, PGE1 tended to increase postsigh apnea incidence and the coefficient of variability (CV) of breathing frequency, suggesting increased respiratory instability. PGE1 also decreased brainstem microglia mRNA and increased neuronal nitric oxide synthase (nNOS) and platelet-derived growth factor-β (PDGF-β) gene expression. Caffeine pretreatment prevented these effects of PGE1, and the adenosine A2A receptor inhibitor MSX-3 had similar preventative effects. Prostaglandin appears to have deleterious effects on brainstem respiratory control regions, possibly involving a microglial-dependent mechanism. The compensatory effects of caffeine or MSX-3 treatment raises the question of whether prostaglandin may also operate on an adenosine-dependent pathway.


Advancing PPG Signal Quality and Know-How Through Knowledge Translation-From Experts to Student and Researcher.
Objective: Despite the vast number of photoplethysmography (PPG) research publications and growing demands for such sensing in Digital and Wearable Health platforms, there appears little published on signal quality expectations for morphological pulse analysis. Aim: to determine a consensus regarding the minimum number of undistorted i.e., diagnostic quality pulses required, as well as a threshold proportion of noisy beats for recording rejection. Approach: Questionnaire distributed to international fellow researchers in skin contact PPG measurements on signal quality expectations and associated factors concerning recording length, expected artifact-free pulses ("diagnostic quality") in a trace, proportion of trace having artifact to justify excluding/repeating measurements, minimum beats required, and number of respiratory cycles. Main Results: 18 (of 26) PPG researchers responded. Modal range estimates considered a 2-min recording time as target for morphological analysis. Respondents expected a recording to have 86-95% of diagnostic quality pulses, at least 11-20 sequential pulses of diagnostic quality and advocated a 26-50% noise threshold for recording rejection. There were broader responses found for the required number of undistorted beats (although a modal range of 51-60 beats for both finger and toe sites was indicated). Significance: For morphological PPG pulse wave analysis recording acceptability was indicated if <50% of beats have artifact and preferably that a minimum of 50 non-distorted PPG pulses are present (with at least 11-20 sequential) to be of diagnostic quality. Estimates from this knowledge transfer exercise should help inform students and researchers as a guide in standards development for PPG study design.


Identification of unique microbiomes associated with harmful algal blooms caused by Alexandrium fundyense and Dinophysis acuminata.
Biotic interactions dominate plankton communities, yet the microbial consortia associated with harmful algal blooms (HABs) have not been well-described. Here, high-throughput amplicon sequencing of ribosomal genes was used to quantify the dynamics of bacterial (16S) and phytoplankton assemblages (18S) associated with blooms and cultures of two harmful algae, Alexandrium fundyense and Dinophysis acuminata. Experiments were performed to assess changes in natural bacterial and phytoplankton communities in response to the filtrate from cultures of these two harmful algae. Analysis of prokaryotic sequences from ecosystems, experiments, and cultures revealed statistically unique bacterial associations with each HAB. The dinoflagellate, Alexandrium, was strongly associated with multiple genera of Flavobacteria including Owenweeksia spp., Maribacter spp., and individuals within the NS5 marine group. While Flavobacteria also dominated Dinophysis-associated communities, the relative abundance of Alteromonadales bacteria strongly co-varied with Dinophysis abundances during blooms and Ulvibacter spp. (Flavobacteriales) and Arenicella spp. (Gammaproteobacteria) were associated with cells in culture. Eukaryotic sequencing facilitated the discovery of the endosymbiotic, parasitic dinoflagellate, Amoebophrya spp., that had not been regionally described but represented up to 17% of sequences during Alexandrium blooms. The presence of Alexandrium in field samples and in experiments significantly altered the relative abundances of bacterial and phytoplankton by both suppressing and promoting different taxa, while this effect was weaker in Dinophysis. Experiments specifically revealed a negative feedback loop during blooms whereby Alexandrium filtrate promoted the abundance of the parasite, Amoebophrya spp. Collectively, this study demonstrates that HABs formed by Alexandrium and Dinophysis harbor unique prokaryotic and eukaryotic microbiomes that are likely to, in turn, influence the dynamics of these HABs.


Evaluation and management of oral lesions in the emergency department.
This article reviews the emergency diagnosis and treatment of oral lesions. It presents a framework for developing a differential diagnosis based on the color of the lesions, describes emergency department interventions, and identifies pitfalls for the emergency physician.


Biological properties of the 2-fluoro-beta-alanine conjugates of cholic acid and chenodeoxycholic acid in the isolated perfused rat liver.
The isolated perfused rat liver was used to examine the hepatic extraction, biliary secretion and effect on bile flow of the 2-fluoro-beta-alanine conjugates of cholic acid and chenodeoxycholic acid. The naturally occurring taurine and glycine conjugates of these bile acids were used for comparisons. The 2-fluoro-beta-alanine conjugates were extracted by the liver to a similar extent as the taurine and glycine conjugates. The biliary secretion rate and increase in bile flow were similar for all the cholic acid conjugates. On the other hand, the maximal biliary secretion rate of the 2-fluoro-beta-alanine conjugate of chenodeoxycholate was similar to that of the glycochenodeoxycholate, but 47% lower than that of taurochenodeoxycholate. In addition, the 2-fluoro-beta-alanine conjugate of chenodeoxycholate produced a decrease in bile flow that was comparable to that observed with the glycochenodeoxycholate (54% vs. 74%), but which was greater than that produced by the taurochenodeoxycholate (12%). In summary, these data demonstrate that the biological properties of the 2-fluoro-beta-alanine conjugates of cholic acid and chenodeoxycholic acid are not markedly different from those of the naturally occurring taurine and glycine conjugates. These data also suggest that the amino acid moiety can influence the biliary secretion and cholestatic properties of chenodeoxycholic acid conjugates.


Proposed bone health screening protocol to identify total knee arthroplasty patients for preoperative DXA.
This study evaluates a novel, simple bone health screening protocol composed of patient sex, age, fracture history, and FRAX risk to identify total knee arthroplasty patients for preoperative DXA. Findings supported effectiveness, with sensitivity of 1.00 (CI 0.92-1.00) and specificity of 0.54 (CI 0.41-0.68) when evaluating for clinical osteoporosis. Bone health optimization is a process where osteoporotic patients are identified, evaluated via modalities such as dual-energy X-ray absorptiometry (DXA), and treated when indicated. There are currently no established guidelines to determine who needs presurgical DXA. This study evaluates the effectiveness of a simple screening protocol to identify TKA patients for preoperative DXA. This prospective cohort study began on September 1, 2019, and included 100 elective TKA patients. Inclusion criteria were ≥ 50 years and primary TKA. All patients obtained routine clinical DXA. The screening protocol defining who should obtain DXA included meeting any of the following: female ≥ 65, male ≥ 70, fracture history after age 50, or FRAX major osteoporotic fracture risk without bone mineral density (BMD) adjustments ≥ 8.4%. Osteoporosis was defined by the World Health Organization (WHO) criteria (T-score ≤ - 2.5) or clinically (T-score ≤ - 2.5, elevated BMD-adjusted FRAX risk, or prior hip/spine fracture). Sensitivity and specificity were calculated. The study included 68 females and 32 males, mean age 67.2 ± 7.7. T-score osteoporosis was observed in 16 patients while 43 had clinical osteoporosis. Screening criteria recommending DXA was met by 69 patients. Screening sensitivity was 1.00 (CI 0.79-1.00) and specificity was 0.37 (CI 0.27-0.48) for identifying patients with T-score osteoporosis. Similar sensitivity of 1.00 (CI 0.92-1.00) and specificity of 0.54 (CI 0.41-0.68) were found for clinical osteoporosis. A simple screening protocol identifies TKA patients with T-score and clinical osteoporosis for preoperative DXA with high sensitivity in this prospective cohort study.


Cannabis use and 'safe' identities in an inner-city school risk environment.
Despite evidence of school effects on drug use, little is known about the social and institutional processes through which these may occur. This study explores how school experiences may shape young people's drug-related attitudes and actions and adds to existing evidence highlighting the importance of drug use in young people's identity construction and group bonding. Case study qualitative data were collected through semi-structured interviews supplemented with observations. Fourteen students aged 14-15 were interviewed in autumn 2006 and again in summer 2007. Five teachers were interviewed. Inner-city secondary schools may constitute risky and insecure environments, and thereby act as structural forces in shaping how students form peer groups and respond to feelings of insecurity. Cannabis use--and identities constructed in relation to this--appeared to play a key role in how students manage insecurity. Black and dual-heritage students formed large, 'safe' ethno-centric school social networks and smoking 'weed' appeared to be an important source of bonding and identity. For some students, a vicious circle may exist whereby the process of 'fitting in' exacerbates difficult and conflicting relationships with teachers and parents, entrenches disengagement from education and leads to further cannabis use. There also appeared to be a diffusion of this 'safe' identity construction beyond these disengaged students. Cannabis and other drug use may also be an important safety strategy and source of bonding for pro-education students who need to be seen 'getting high' as well as 'aiming high'. While some students constructed 'safe' and 'sweet' identities others were constrained by their position and resources at school. Inner-city schools may both reflect and reproduce existing patterns of drug use. The concept of risk hierarchies may be important when designing and evaluating school-based drug-prevention strategies.


Stereological analysis of astrocyte and microglia in aging mouse hippocampus.
Recent evidence suggests neuroglia-mediated inflammatory mechanisms may stimulate neurodegenerative processes in mammalian brain during aging. To test the hypothesis that the number of microglia and astrocytes increase in the hippocampus during normal aging, unbiased stereological techniques were used to estimate total cell number in hippocampal subregions (CA1, dentate gyrus and hilus) of male C57BL/6J mice of different ages: 4-5 months, 13-14 months and 27-28 months. Immunocytochemical visualization for microglia and astrocytes were via Mac-1 and GFAP antibody, respectively. Estimates of total microglia and astrocyte number were assessed using the optical fractionator. No statistically significant age differences were found in the numbers of microglia or astrocytes in the hippocampal regions sampled. These findings suggest that age-related increases in the total numbers of hippocampal microglia and astrocytes is not causal for observed age-related increases in cytokine response.


Feet, bridges, and pillars in triad junctions of mammalian skeletal muscle: their possible relationship to calcium buffers in terminal cisternae and T-tubules and to excitation-contraction coupling.
The structure of the triad junction was examined in thin sections of mammalian fast-twitch skeletal muscle. The aims of the experiments were twofold: first, to examine relationships between the contents of the junctional gap and the terminal cisternae that could be significant in excitation-contraction coupling and, second, to look for structures in the transverse tubules that could support a calcium buffer system. Procedures known to stabilize cytoskeletal elements were used in an attempt to retain the original structure. "Feet," "pillars" and "bridges" were often seen side by side in the same junction. In one such junction, the average center-to-center spacing between four bridges was 30.9 +/- 1.7 nm and between five foot-like structures was 29.2 +/- 1.4 nm. The subunit structure of the feet could be seen in many sections. The lumen of the terminal cisternae was filled with a tetragonal network of calsequestrin which formed parallel strands near the junctional membrane, in register with the feet. The strands overlay the area occupied by "rods" seen in freeze-fracture replicas of terminal cisterna membrane. The contents of the transverse tubules were aggregated into bands, or "tethers," which extended across the short axis of the tubule at regular intervals of about 30 nm. The tethers consisted of flattened discs, stacked across the long axis of the tubule, aligned with the junctional feet. Lanthanum staining of the tethers indicated cationic binding sites that could buffer luminal calcium ion concentration in the vicinity of the voltage sensor for contraction. It is suggested (i) that the control of calcium concentration near the voltage sensor is necessary for normal activation, (ii) that feet, pillars and bridges are different images of a spanning structure, and (iii) that the regular alignment of tethers, feet and calsequestrin is functionally significant in excitation-contraction coupling.


Performance experimental data of a polymer electrolyte fuel cell considering the variation of the relative humidity of reactants gases.
The data collected in this article is based on a performance test of a polymer electrolyte fuel cell (PEFC). The behavior the different parameters of a PEFC is analyzed considering different aspects relative to the inlet gases temperatures. The fuel cell was evaluated by means of a current sweep at different percentages of relative humidity between the feed gas and the cell. The relative humidity values were established by means of the temperature setting. The data presented show the experimental response of the cell in real time, which can be used to perform a depth analysis or they can be a starting point for material and performance investigation. In addition, charts presenting the voltage and power density behavior as a function of the volumetric flows of the anode (H2) as well as cathode (O2). The data presented in this article are originally from our research performed in [1].


Synthesis and characterization of Ni-doped anatase TiO2 loaded on magnetic activated carbon for rapidly removing triphenylmethane dyes.
In this work, we employed the in situ synthesis method to implant Fe3O4 into activated carbon (AC), in which the synthesis of the magnetic AC (MAC) was realized. Thence, Ni-doped anatase TiO2 (NATiO2) were anchored on different addition amount of MAC to synthesize the series of Ni-TiO2/MAC photocatalysts. The chemical compositions and physical properties of these nanocomposites were analyzed by various characterization technologies. The photocatalytic capabilities of as-produced materials were then investigated via adsorption and photodegradation of triphenylmethane dyes (TPMs) as crystal violet (CV), basic fuchsine (BF), and malachite green (MG) solution. The results revealed that the removal of Ni-TiO2/AC, Ni-TiO2/2MAC, Ni-TiO2/4MAC, and Ni-TiO2/8MAC on TPMs is a very fast process and the removal efficiency can almost reach to about 90% in 10 min, and the catalyst has good cycle stability and is easy to be reused. This work provides a novel, low-cost, and effective way to rationally design and synthesize TiO2-based photocatalysts for effective removal of TPMs.


Cellular aspects of breast cancer: workshop report.
It has not been possible in this review to cover all the submitted posters nor indeed all the points discussed during the workshop session. It is hoped, however, that major points of interest have been presented and that areas where there is consensus and areas where controversy remains concerning the clinical value of particular investigations have been highlighted. It is also hoped that some new potential growth areas for investigation of tumour biology have been defined.


A T1083C polymorphism in the human adenosine A2a receptor gene is not associated with essential hypertension.
The adenosine A2a receptor (A2aAR) gene is thought to be involved in essential hypertension because adenosine elicits vasodilation and decreases arterial blood pressure via this receptor, and because disruption of the A2aAR gene increases blood pressure in mice. Therefore, using a restriction fragment length polymorphism (RFLP) of the A2aAR gene, we performed an association study in patients with essential hypertension. One hundred forty-two patients with essential hypertension and 142 age-matched subjects with normal blood pressure were studied. Polymerase chain reaction (PCR) was applied to amplify the T1083C polymorphic site in the A2aAR gene, and restriction analysis of the PCR product was employed to score the T and C alleles. Overall distributions of allele frequencies in the two groups were not significantly different. Thus, the alleles detected by this RFLP polymorphism in the A2aAR gene are not associated with essential hypertension.


The Effect of Bifidobacterium on Reducing Symptomatic Abdominal Pain in Patients with Irritable Bowel Syndrome: A Systematic Review.
Probiotics, specifically Bifidobacteria, may improve abdominal pain in patients with irritable bowel syndrome (IBS); however, results from randomised controlled trials (RCTs) are conflicting. Here, we systematically reviewed the efficacy of Bifidobacteria on abdominal pain in IBS. We searched MEDLINE, EMBASE and the Cochrane Controlled Trials Register from inception to 20 May 2019, without language or date restrictions. The search strategy comprised of the combination of three concepts: supplementation, abdominal pain, and IBS. Inclusion criteria included double-blind placebo-controlled RCTs featuring Bifidobacteria supplementation in Rome-diagnosed IBS patients. A total of 8 RCTs involving a total of 1045 patients with Rome diagnosed IBS were included. The dose of total Bifidobacteria ranged from 106 to > 1011 cfu (colony-forming unit) and duration of supplementation ranged between 2 and 8 weeks. Bifidobacteria was delivered through either intake of fermented milk products, encapsulation or via a malted milk beverage, with all studies assessing abdominal pain via a visual analogue Likert scale. From the studies included, 50% (n = 4) of studies found a statistically significant improvement in abdominal pain following Bifidobacteria supplementation compared to placebo, 38% (n = 3) of studies found non-significant improvements and 12% (n = 1) showed a statistically significant dose-response effect of improvement. The evidence shows a heterogeneity of effect for Bifidobacteria dependent upon strain, dosage and delivery method. While not all studies demonstrate significant improvements in abdominal pain, none of the selected studies reported an increase in pain or other adverse effects.


Acute genetic perturbation of exocyst function in the rat calyx of Held impedes structural maturation, but spares synaptic transmission.
The exocyst is an octameric protein complex mediating polarized secretion by tethering vesicles to target membranes. In non-vertebrate neurons, the exocyst has been associated with constitutive membrane addition at growth cones and nerve terminals, but its function in synaptic vesicle trafficking at mammalian nerve terminals remains unclear. Here, we examined the role of the exocyst complex in immature postnatal day (P)13 and mature P21 rat calyces of Held. Exo70, an exocyst subunit conferring membrane anchoring of the complex, was tagged with green fluorescent protein (GFP) and overexpressed as a full-length subunit or as a dominant-negative C-terminally truncated variant (Exo70ΔC) disrupting membrane targeting. In vivo expression of the Exo70 subunits in the calyx was achieved by stereotaxic adeno-associated virus-mediated gene transfer into globular bushy cells of the rat ventral cochlear nucleus at P2. Overexpression of dominant-negative Exo70ΔC, but not full-length Exo70, decreased the structural complexity and volume of calyces, as assayed by confocal microscopy and three-dimensional reconstructions. The distribution of active zones and synaptic vesicles remained unaffected. Neither perturbation changed the characteristics of spontaneous and evoked neurotransmitter release, short-term depression or recovery from depression. Together, these data suggest that in central mammalian synapses, the exocyst complex mediates the addition of membrane during postnatal presynaptic maturation, but does not function as a tethering complex in local recycling of vesicles within the synaptic vesicle cycle.


Peptidomics analysis of enzymatic hydrolysis beef.
In this study, we investigated the changes in the composition of peptides during the digestion of tenderized beef treated with commercial proteinase K, flavourzyme, and bromelain. The degree of hydrolysis (DH) values before and after simulating gastric digestion were highest with proteinase K treatment. In the proteinase K-treated sample, the highest number of missing peptides was identified after gastrointestinal digestion. Additionally, the maximum number of new peptides was identified during gastric digestion. The flavourzyme is the only exopeptidase among the three enzymes, and the sample treated with it could produce more unique peptides after gastrointestinal digestion. Enzymatic tenderization altered the peptide composition and bioactivity of beef proteins during gastrointestinal digestion. The number of peptides, as well as unique peptides in the protease-treated sample, were more than those in control through gastric digestion. In contrast, the opposite was observed post gastrointestinal digestion. The online version contains supplementary material available at 10.1007/s10068-022-01122-y.


[4,4'-D-diaminopropionic acid]gramicidin S: a synthetic gramicidin S analog with antimicrobial activity against Gram-negative bacteria.
Gramicidin S is especially active against Gram-positive bacteria; e.g., Staphylococcus aureus. An analog, [4,4'-D-diaminopropionic acid]gramicidin S, which contains D-diaminopropionic acid residues instead of D-phenylalanine residues, has been synthesized. This analog is active against some of the Gram-negative bacteria; e.g., Escherichia coli and Salmonella typhosa. Activities of several related analogs are discussed.


Knuckle pads in Dupuytren's disease.
Knuckle pads are not exclusively found in persons with Dupuytren's disease, but they occur four times more often in these patients than in the general population (in Norway). Occurrence of knuckle pads in persons with Duputyren's disease does not seem to imply a higher predisposition or a more aggressive disease.


Solvent states and spectroscopy of doped helium clusters as a quantum-chemistry-like problem.
The Full-Configuration-Interaction Nuclear-Orbital (FCI-NO) approach [J. Chem. Phys., 2009, 131, 19401], as the implementation of the quantum-chemistry ansatz, is overviewed and applied to (He)N-Cl2(X) clusters (N≤ 4). The ground and excited states of both fermionic (3)He and bosonic (4)He [see also, J. Phys. Chem. Lett., 2012, 2, 2145] clusters are studied. It is shown that the FCI-NO approach allows us to overcome three main difficulties: (1) the Fermi-Dirac (Bose-Einstein) nuclear statistics; (2) the wide (highly anharmonic) amplitudes of the He-dopant and He-He motions; and (3) both the weakly attractive (long-range) and the strongly repulsive (short-range) interaction between the helium atoms. Special emphasis is placed on the dependence of the cluster properties on the number of helium atoms, and on the comparison between the two helium isotopes. In particular, we analyze the analogies between quantum rings comprising electrons and (3)He atoms. The synthetic vibro-rotational Raman spectra of Cl2(X) immersed in ((3,4)He)N clusters (N≤ 4) are discussed as a function of the cluster size and the nuclear statistics. It is shown that the Coriolis couplings play a key role in modifying the spectral dopant profile in (3)He. Finally, we point out possible directions for future research using the quantum-chemistry ansatz.


Lignocellulosic fibres surface interactions in enzymatic reaction using data-mining.
Lignocellulosic fibres modification focused so far essentially on the resulting material properties to create functional fibres instead of determining the reaction influencing parameters. Using a data-mining algorithm, surface chemical composition of the fibres after modifications was compared to multiple signals. A 24 h reaction at either 25 °C or 60 °C, pH5 was conducted in presence of trans-ferulic acid, laccase, and lignocellulosic fibres (flax, hemp, or cellulose) having different chemical surface composition. Dimers and trimers were detected in variable concentrations in the reaction filtrate and extractive. At 25 °C, crystalline cellulose, amorphous cellulose, xylans, mannans, and lignins were well correlated to specific reaction products while at 60 °C, only lignins and xylan were found correlated to reaction products. Fibres surface composition affected the extractive profile. Lignocellulosic surface composition influence on the product formed was unveiled using a data mining approach. This study presents a way to unveil non-evident chemical interface interaction in reactions.


Patient satisfaction with different asthma-training variants.
Information on patient satisfaction with professional health care delivery in asthmatics is rare, and the question as to how asthma education programmes affect such satisfaction has not yet been addressed. This multi-centre study investigated three different variants of an asthma education programme for adults. Patients participated either in variant B (basic training with teacher-directed presentation, two 90 min sessions), variant C (comprehensive training, four sessions), variant D (including additional psychological components, minimum five sessions), or variant E (equivalent to B in outpatient context). At the end of the training the perceived satisfaction with the programme and reported personal benefit were assessed by means of a new inventory that was developed. The sample comprised N = 320 patients (n = 244 inpatients and n = 76 outpatients) ranging in age from 18 to 80 years (M = 46.3 years). The average total score for educational satisfaction of all participants was 8.0 (maximum score = 10). Choice of topics was judged particularly positive (M = 8.58), followed by the structure of training/setting-conditions (M = 8.0). The perceived personal benefit from the training was evaluated less positively (M = 7.32). When comparing the programme variants, patients' average satisfaction tends to increase with the extent and intensity of the training offered. Outpatients showed significantly less satisfaction in contrast to inpatients. Inpatients' satisfaction with the three variants increased with extensiveness and intensity of the training, while the contrast of between-group comparisons (B-C, B-D and C-D) was significant. Assessment of the patient's view of health care services should complement standardised evaluation methods, especially in multi-intervention rehabilitation programmes. Rehabilitation-specific diagnostic measures should be developed and validated in order to better assess patients' satisfaction and training efficacy from the patient's perspective. Further study is recommended on how far individualised training measures might increase subjective benefit.


Heat over heritability: Increasing body size in response to global warming is not stabilized by genetic effects in Bechstein's bats.
How well populations can cope with global warming will often depend on the evolutionary potential and plasticity of their temperature-sensitive, fitness-relevant traits. In Bechstein's bats (Myotis bechsteinii), body size has increased over the last decades in response to warmer summers. If this trend continues it may threaten populations as larger females exhibit higher mortality. To assess the evolutionary potential of body size, we applied a Bayesian 'animal model' to estimate additive genetic variance, heritability and evolvability of body size, based on a 25-year pedigree of 332 wild females. Both heritability and additive genetic variance were reduced in hot summers compared to average and cold summers, while evolvability of body size was generally low. This suggests that the observed increase in body size was mostly driven by phenotypic plasticity. Thus, if warm summers continue to become more frequent, body size likely increases further and the resulting fitness loss could threaten populations.


Long-term evaluation of nerve regeneration in a biodegradable nerve guide.
Nerve regeneration using artificial biodegradable conduits is of increasing interest. The aim of this study is to evaluate the regeneration and maturation of a nerve after long-term implantation (2 years) of a biodegradable poly-L-lactide/poly-epsilon-caprolactone (PLLA/PCL) copolymeric nerve guide in the sciatic nerve of the rat. After harvesting, we evaluated both the regenerated nerves and the controls, using light microscopy, transmission electron microscopy, and morphometric techniques. Remnants of biomaterial were still present after 2 years of implantation, but the foreign body reaction was very mild at this stage, due to the rounded shapes of the polymer debris. Morphometric analysis showed significant differences between the regenerated nerve and the normal sciatic nerve: the number of myelinated fibers is higher, and the mean fiber diameter of the myelinated fibers in the regenerated nerve is smaller. In conclusion, the results demonstrate that the new PLLA/PCL nerve guide can provide optimal conditions for regeneration and maturation of damaged nerves.


Neighborhood Crime and Perception of Safety as Predictors of Victimization and Offending Among Youth: A Call for Macro-Level Prevention and Intervention Models.
This paper is one of two in a series that reports detailed findings from a larger study that simultaneously explored individual, family and neighborhood level predictors of victimization and offending among youth. The current analysis aims to identify which neighborhood level factors have better predictive power with regard to type of victimization (direct and vicarious measures) and total offending overtime (Wave 1 and Wave 2). METHODS: Path analysis was conducted using data from a multi-wave, panel study (N=625) of youth ages 16-19 at Wave 1. A best fitting model was determined showing causal pathways from neighborhood level factors including crime and perception of safety, to direct and vicarious victimization through exposure to violence, and subsequent offending. FINDINGS: Neighborhood crime significantly predicted property victimization. Neighborhood crime and perception of safety significantly predicted vicarious victimization by exposure to violence in the neighborhood. Neighborhood crime and perception of safety were significantly associated with Wave 1 offending. Findings highlight the need for professionals who work with youth to be cognizant of how their environments influence their lives. Prevention and intervention models seeking to create sustainable change among youth should consider mezzo and macro level components that build and strengthen neighborhood capacity through community partnerships.


Relationships among the genera Ashbya, Eremothecium, Holleya and Nematospora determined from rDNA sequence divergence.
Species of the genera Ashbya, Eremothecium, Holleya, and Nematospora were compared from extent of divergence in a 580-nucleotide region near the 5' end of the large subunit (26S) ribosomal DNA gene. The four genera are closely related and comprise a subclade of the hemiascomycetes. Because the taxa show little divergence, it is proposed that all be placed in the genus Eremothecium. The family Eremotheciaceae, fam. nov., is proposed.


Antiglaucoma drugs: The role of preservative-free formulations.
Hypersensitive reactions to eyedrops are a common finding in clinical practice and represent a frequent cause of discontinuation of the therapy. Moreover, experimental and clinical studies show that long term use of topical drugs may induce ocular surface changes causing discomfort and potentially negatively affecting the compliance to the treatment as well as the success rate of filtering procedures. The exact mechanism involved and the roles of the active compound and the preservatives in inducing such detrimental effects of ophthalmic solutions are unclear. During the last years several antiglaucoma agents have been marketed as either preservative-free or benzalkonium chloride-free formulations in an attempt to reduce the adverse effects related to preservatives. This paper summarizes the body of evidence from existing studies about preservatives in antiglaucoma eyedrops, focusing on the latest compounds commercially available. A systematic review of the literature was performed. Current research is focusing not only on the efficacy of the drugs but also on their tolerability. Based on the existing data, there is a rationale to support the use of benzalkonium-free solutions whenever possible, especially in patients suffering from concomitant ocular surface diseases, experiencing local side effects and in those expected to need multiple and prolonged topical treatments.


A prospective study of multivitamin supplement use and risk of breast cancer.
The authors evaluated the association between multivitamin supplement use and breast cancer risk in a completed trial. At baseline (1992-1995), 37,920 US women aged > or =45 years and free of cancer provided detailed information on multivitamin supplement use. During an average of 10 years of follow-up, 1,171 cases of invasive breast cancer were documented. Multivitamin use was not significantly associated with overall risk of breast cancer. Compared with the risk for never users, the multivariable relative risks were 0.97 (95% confidence interval: 0.81, 1.16) for past users and 0.99 (95% confidence interval: 0.82, 1.19) for current users. Current multivitamin use for > or =20 years or > or =6 times/week was also not significantly associated with risk. Multivitamin use was nonsignificantly inversely associated with risk of breast cancer among women consuming > or =10 g/day of alcohol and with risk of estrogen receptor negative-progesterone receptor negative breast cancer. Multivitamin use was nonsignificantly associated with a reduced risk of developing < or =2-cm breast tumors but an increased risk of >2-cm tumors. The authors' data indicate no overall association between multivitamin use and breast cancer risk but suggest that multivitamin use might reduce risk for women consuming alcohol or decrease risk of estrogen receptor negative-progesterone receptor negative breast cancer.


Screening for light-for-gestational age infants: a comparison of three simple measurements.
Fundal height, abdominal girth and abdominal length were measured during routine examination in our antenatal clinic. Standard graphs of fundal height, abdominal girth and the ratio of fundal height to abdominal length were drawn using data from 250 consecutive women with uncomplicated pregnancies. These graphs were used to predict low-birthweight-for-gestation (LGA) in 150 high-risk and 208 low-risk pregnancies. Fundal height measurements had high sensitivity (78-84%) and specificity (84-88%) and were therefore most reliable in screening for LGA babies.


Isolated episode of exercise-related ventricular fibrillation in a healthy athlete.
A 19-year-old man had a documented ventricular fibrillation during a football match. He was a top-level athlete, and after successful resuscitation, he was extensively studied. Electrolytic disturbances, long QT syndrome and nervous mechanisms were all ruled out. Right and left ventriculography, coronary angiograms, electrophysiologic study and endomyocardial biopsy all proved normal. A beta-blocker was empirically given in the absence of any reliable test to guide therapy more scientifically.


Perinatal death investigations: What is current practice?
Perinatal death (PD) is a devastating obstetric complication. Determination of cause of death helps in understanding why and how it occurs, and it is an indispensable aid to parents wanting to understand why their baby died and to determine the recurrence risk and management in subsequent pregnancy. Consequently, a perinatal death requires adequate diagnostic investigation. An important first step in the analysis of PD is to identify the case circumstances, including relevant details regarding maternal history, obstetric history and current pregnancy (complications are evaluated and recorded). In the next step, placental examination is suggested in all cases, together with molecular cytogenetic evaluation and fetal autopsy. Investigation for fetal-maternal hemorrhage by Kleihauer is also recommended as standard. In cases where parents do not consent to autopsy, alternative approaches such as minimally invasive postmortem examination, postmortem magnetic resonance imaging, and fetal photographs are good alternatives. After all investigations have been performed it is important to combine findings from the clinical review and investigations together, to identify the most probable cause of death and counsel the parents regarding their loss.


New bisbenzylisoquinoline alkaloids from Cocculus laurifolius.
Two new bisbenzylisoquinoline alkaloids, alpha, alpha'-dioxo-7'-O-demethylstebisimine (1) and 7'-O-demethylstebisimine (2), along with 14 known alkaloids, were isolated from the roots of Cocculus laurifolius. These alkaloids were characterized mainly by spectroscopic methods.


The Adolescent Experience of Filling Prescriptions for the Treatment of Sexually Transmitted Infections.
To describe adolescents' experience with treatment adherence when prescribed antibiotics for sexually transmitted infections (STIs) in emergency departments (ED). Adolescents diagnosed with STIs in two EDs and prescribed outpatient treatment were interviewed to identify barriers and facilitators to treatment adherence. Of 26 participants, 46.2% filled their prescriptions, 38.5% returned to the ED for treatment, 7.7% received treatment elsewhere, and 7.7% received no treatment. All adolescents who filled their prescriptions notified caregivers of their diagnosis, compared with 50.0% who returned to the ED (P = 0.01). Adolescents identified cost, transportation, lack of insurance card, and lack of knowledge as perceived barriers to treatment adherence. Less than half of the participants were interested in school-based health clinics due to anonymity concerns. Most expressed interest in using mobile health (mHealth) to overcome adherence challenges. Adolescents identified multiple barriers to treatment adherence. Future work should explore the role of caregiver communication and incorporation of mHealth.


Postoperative outcomes of initial varus versus initial valgus proximal humerus fracture: A systematic review and meta-analysis.
Our objective was to perform a systematic review of the literature and conduct a meta-analysis to investigate the effect of initial varus or valgus displacement of proximal humerus on the outcomes of patients with proximal humerus fractures treated with open reduction and internal fixation. In accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement standards, we performed a systematic review. Electronic databases MEDLINE, EMBASE, CINAHL and the Cochrane Central Register of Controlled Trials (CENTRAL) were searched to identify randomised and non-randomised studies comparing postoperative outcomes associated with initial varus versus initial valgus displacement of proximal humerus fracture. The Newcastle-Ottawa scale was used to assess the methodological quality and risk of bias of the selected studies. Fixed-effect or random-effects models were applied to calculate pooled outcome data. We identified two retrospective cohort studies and one retrospective analysis of a prospective database, enrolling a total of 243 patients with proximal humerus fractures. Our analysis showed that initial varus displacement was associated with a higher risk of overall complication (RR 2.28, 95% CI 1.12-4.64, P = 0.02), screw penetration (RR 2.30, 95% CI 1.06-5.02, P = 0.04), varus displacement (RR 4.38, 95% CI 2.22-8.65, P < 0.0001), and reoperation (RR 3.01, 95% CI 1.80-5.03, P < 0.0001) compared to valgus displacement. There was no significant difference in avascular necrosis (RR 1.43, 95% CI 0.62-3.27, P = 0.40), infection (RR 1.49, 95% CI 0.46-4.84, P = 0.51), and non-union or malunion (RR 1.37, 95% CI 0.37-5.04, P = 0.64). The best available evidence demonstrates that initial varus displacement of proximal humerus fractures is associated with higher risk of overall complication, screw penetration, varus displacement, and reoperation compared to initial valgus displacement. The best available evidence is not adequately robust to make definitive conclusions. Further high quality studies, that are adequately powered, are required to investigate the outcomes of initial varus and valgus displacement in specific fracture types. Level II.


Plasmodium vivax genetic diversity: microsatellite length matters.
The Plasmodium vivax genome is very diverse but has a relatively low abundance of microsatellites. Leclerc et al. had shown that these di-nucleotide repeats have a low level of polymorphism, suggesting a recent bottleneck event in the evolutionary history of P. vivax. By contrast, in a recent paper, Imwong et al. show that there is a very high level of microsatellite diversity. The difference in these results is probably due to the set array lengths chosen by each group. Longer arrays are more diverse than are shorter ones because slippage mutations become exponentially more common with an increase in array length. These studies highlight the need to consider carefully the application and design of studies involving microsatellites.


Determination of the number of detergent molecules associated with the reaction center protein isolated from the photosynthetic bacterium Rhodopseudomonas viridis. Effects of the amphiphilic molecule 1,2,3-heptanetriol.
Detergent-free reaction center (RC) proteins from the photosynthetic bacterium Rhodopseudomonas viridis were obtained using Bio-Beads SM-2. With these RCs, the amount of detergent molecules associated with the protein was measured by determining the detergent concentration at which re-solubilization occurred as a function of the RC concentration. For N,N-dimethyl dodecylamine-N-oxide (LDAO), Triton X-100 and beta-octylglucoside 260 +/- 30,105 +/- 10 and 360 +/- 100 detergent molecules were necessary to dissolve the protein, respectively. With this technique we have studied the effect of the amphiphilic molecule 1,2,3-heptanetriol, which is essential in the crystallization process of these RCs. Addition of 5% 1,2,3-heptanetriol reduces the value for LDAO to 120 +/- 20 LDAO/RC, supporting the notion that crystallization of the RCs is promoted by increasing the number of protein-protein contacts.


Heating temperature dependence of molecular characteristics and biological response for biomass pyrolysis volatile-derived water-dissolved organic matter.
The utilization of biomass pyrolysis volatile-derived water-dissolved organic matter (WOM, often called wood vinegar) determines sustainable recycling of biomass. Further, pyrolysis temperature significantly controls the cracking of biomass components, resulting in various molecular compositions and biological responses of WOM. Although it has been widely used in the agriculture, the relationship between molecular compositions and biological responses affected by heating temperature is still unclear. Here, it was observed that the WOM concentration increased with increasing temperatures and the pyrolysis of 1 g biomass can generate ~ WOM with 36.24 mg C. Moreover, with increasing pyrolysis temperatures, the generated WOM consisted of more phenols but fewer alcohols, furans, acids, and ketones, and demonstrated characteristics of higher aromaticity and lower m/z molecular weight. Due to the enhanced polarity, high temperatures promoted the solubility of WOM. Germination tests show that low pyrolysis temperatures-derived WOM (< 400 °C) with large-molecular-weight and low oxygen-containing (low O/Cwa) promoted plant growth, while high temperatures-derived WOM (> 400 °C) with small-molecular-weight and high oxygen-containing (high O/Cwa) inhibited growth. These results suggest that WOM can be separately collected at different pyrolysis temperatures to achieve sustainable recycling of pyrolysis volatile.


Treatment of neuropathic cancer pain with continuous intrathecal administration of S +-ketamine.
The effective treatment of patients suffering from neuropathic cancer pain remains a clinical challenge. When patients experience either insufficient analgesia or problematic side-effects after opioid administration, intrathecal administration of morphine and other medications such as bupivacaine and clonidine may offer significant advantages. Additionally, ketamine, a non-competitive N-methyl-D-Aspartate-receptor antagonist is able to alter pain perception at the spinal level. Because of the potential neurotoxicity after neuraxial use of racemic ketamine, intrathecal administration of the preservative-free active compound, S (+)-ketamine may be a valuable alternative. In this paper, we present a patient with severe neuropathic cancer pain successfully treated by continuous intrathecal infusion of morphine, bupivacaine, clonidine and S (+)-ketamine. Moreover, quality of life measurements before and 3 weeks after the start of spinal treatment revealed an improvement in pain relief and a higher overall quality of life. No clinical signs of neurologic deficit were observed during spinal treatment with S (+)-ketamine. However, the continuous intrathecal administration of S (+)-ketamine should be considered as the last resort because there are no preclinical safety data with relevant concentrations on intrathecal use of S (+)-ketamine.


Early fire detection for underground diesel fuel storage areas.
With the increased use of mobile diesel-powered equipment in underground mines, the fire risk posed by underground diesel fuel storage areas is a concern. To reduce the risk associated with the storage and transfer of large quantities of diesel fuel in permanent underground mine storage areas, an experimental study was conducted to investigate the responses of different sensors for early detection of diesel fuel fires in a storage area. Fire sensors tested in this study were four carbon monoxide (CO) sensors, two smoke sensors, and one flame sensor. A series of fire tests were conducted in the NIOSH Safety Research Coal Mine, Bruceton, PA, using various fire sizes at different ventilation airflow velocities and fire locations. Response times for different sensors were analyzed, and the results suggest that the flame sensor and smoke sensors resulted in shorter response times in most tests compared to the CO sensors. Based on the test results, the appropriate sensor locations for early fire detection in a diesel fuel storage area were identified. The results of this study can help mining companies to select appropriate fire sensors for underground diesel fuel storage areas and improve the deployment of these sensors to ensure the safety of underground miners.


Trends in hospital treatments for peripheral arterial disease in the United States and association between payer status and quality of care/outcomes, 2007-2011.
This study sought to identify the temporal trends of presenting diagnoses and vascular procedures performed for peripheral arterial disease (PAD) along with the rates of procedures and in-hospital outcomes by payer status. Previous studies suggest that patients with Medicare, Medicaid, or lack of insurance receive poorer quality of care leading to worse outcomes. We analyzed 196,461,055 discharge records to identify all hospitalized patients with PAD records (n=1,687,724) from January 2007 through December 2011 in the Nationwide Inpatient Sample database. The annual frequency of vascular procedures remained unchanged during the study period. Patients with Medicaid were more likely to present with gangrenes, whereas patients with Medicare were more likely to present with ulcers. After adjustment, patients with Medicare and Medicaid were more likely to undergo amputations when compared with private insurance/HMO (OR=1.13, 95% CI=1.10-1.16 and OR=1.24, 95% CI=1.20-1.29, respectively). Patients with both Medicare and Medicaid were less likely to undergo bypass surgery (OR=0.82, 95% CI=0.81-0.84 and OR=0.87, 95% CI=0.85-0.90, respectively), but more likely to undergo endovascular procedures (OR=1.18, 95% CI=1.17-1.20 and OR=1.03, 95% CI=1.01-1.06, respectively). Medicare and Medicaid status versus private insurance/HMO was associated with worse adjusted odds of in-hospital outcomes, including mortality after amputations, endovascular procedures, and bypass surgeries. In this analysis, patients with Medicare and Medicaid had more comorbid conditions at baseline when compared with private insurance/HMO cohorts, were more likely to present with advanced stages of PAD, undergo amputations, and develop in-hospital complications. These data unveil a critical gap and an opportunity for quality improvement in the elderly and those with poor socioeconomic status.


Inactivation of a gene for a fibronectin-binding protein of the oral bacterium Streptococcus mutans partially impairs its adherence to fibronectin.
A sequence of 1647 base pairs in length of Streptococcus mutans DNA that encodes for a 63-kDa protein with significant amino acid similarity with fibronectin-binding proteins of Streptococcus pyogenes and Streptococcus gordonii was cloned. The putative recombinant fibronectin-binding protein of S. mutans was purified using affinity chromatography and the cloned protein was used to prepare polyclonal antibodies against the recombinant protein. In immunoblot assays, antibodies against the S. pyogenes fibronectin-binding protein, FBP54, were cross-reactive with the S. mutans protein that was designated SmFnB. Additionally, antibodies to the S. mutans SmFnB protein reacted with the S. pyogenes FBP54 protein. The S. mutans SmFnB protein was found to bind to immobilized fibronectin in a concentration dependant manner. A mutant strain of S. mutans M51 that was constructed by allelic exchange did not express the SmFnB protein. This mutant strain, S. mutans DeltaSmFnB, was determined in an ELISA to bind to immobilized fibronectin 30% less when compared to the parental strain S. mutans M51. The results are consistent with the conclusion that the 63-kDa SmFnB protein of S. mutans is a fibronectin-binding protein that may contribute to the interaction of S. mutans with damaged heart tissue during pathogenesis of infective endocarditis. Also, the study suggests that multiple molecules may mediate the interaction of S. mutans with fibronectin.


Biochemical conservation of recombinant Drosophila tyrosine hydroxylase with its mammalian cognates.
Dopamine modulates several behavioral and developmental events; in the fruit fly Drosophila melanogaster, dopamine is a neurotransmitter, a neuromodulator, and a developmental signal. Studies in mammals suggest that these diverse roles for dopamine have been evolutionarily conserved. Fundamental regulation of dopamine occurs via tyrosine hydroxylase (TH), the first and rate-limiting enzyme in the catecholamine biosynthetic pathway. Mammalian TH is acutely regulated via phosphorylation-dephosphorylation mechanisms, which occur as a direct consequence of nerve stimulation. We have shown that the Drosophila homolog of TH, DTH, shares over 50% sequence identity with mammalian TH, and the serine residue corresponding to the major site of phosphorylation is conserved. We demonstrate using recombinant DTH protein generated in E. coli that its regulatory biochemical mechanisms closely parallel those from mammals. Drosophila thus provides a highly conserved and tractable model system in which to test the functional consequences of perturbing TH activity by acute regulatory mechanisms.


Treatment of a nephrotic syndrome by endoscopic removal of a villous adenoma of the duodenum.
We report the case of a patient diagnosed with a villous adenoma of the duodenum showing high degree dysplasia who developed a nephrotic syndrome (NS) due to a membranous nephropathy (MN), demonstrated by renal biopsy. Only the endoscopic resection of the duodenal adenoma could control the NS. The first manifestation of a MN is often the development of a NS. Up to 20% of patients older than 65 years who develop a MN have cancer. Tumours most often identified are those of lung, prostate and digestive tract. A renal biopsy is required to identify this type of nephropathy. If a diagnosis of MN is made, an associated tumour should be looked for.


Bifidobacterium adolescentis orchestrates CD143+ cancer-associated fibroblasts to suppress colorectal tumorigenesis by Wnt signaling-regulated GAS1.
The interplay between gut microbiota and tumor microenvironment (TME) in the pathogenesis of colorectal cancer (CRC) is not well explored. Here, we elucidated the functional role of Bifidobacterium adolescentis (B.a) on CRC and investigated its possible mechanism on the manipulation of cancer-associated fibroblasts (CAFs) in CRC. Different CRC animal models and various cell line models were established to explore the function of B.a on CRC. The single-cell RNA sequencing (scRNA-seq) or flow cytometry was used to detect the cell subsets in the TME of CRC. Western blot, quantitative real-time polymerase chain reaction (qRT-PCR), or immunofluorescence staining were performed to examine the activation of Wnt signaling and growth arrest specific 1 (GAS1) on CD143+ CAFs. Chromatin immunoprecipitation quantitative real-time PCR (CHIP-qPCR) was performed to investigate the regulation of transcription factor 4 (TCF4) on GAS1. Multi-immunofluorescence assay examined the expression level of CD143 and GAS1 on tissue microarray. We found that B.a abundance was significantly reduced in CRC patients from two independent cohorts and the bacteria database of GMrepo. Supplementation with B.a suppressed ApcMin/+ spontaneous or AOM/DSS-induced tumorigenesis in mice. scRNA-seq revealed that B.a facilitated a subset of CD143+ CAFs by inhibiting the infiltration of Th2 cells, while promoting the TNF-alpha+ B cells in TME. CD143+ CAFs highly expressed GAS1 and exhibited tumor suppressive effect. Mechanistically, GAS1 was activated by the Wnt/β-catenin signaling in CD143+ CAFs. B.a abundance was correlated with the expression level of CD143 and GAS1. The level of CD143+ CAFs predicted the better survival outcome in CRC patients. These results highlighted that B.a induced a new subset of CD143+ CAFs by Wnt signaling-regulated GAS1 to suppress tumorigenesis and provided a novel therapeutic target for probiotic-based modulation of TME in CRC.


Arthroscopically assisted anatomical coracoclavicular ligament reconstruction using tendon graft.
We describe a method of arthroscopically assisted, mini-open, anatomical reconstruction of the coracoclavicular ligament. This method restores both components of the native ligament with the aim of achieving maximum stability with minimal disruption of the normal anatomy. Using the same principles of ligament reconstruction that are employed in other joints, transosseous tunnels are created following the native footprints of the conoid and trapezoid ligaments and an autologous graft is fixed using a PEEK screw. Adequate healing of the ligament occurs within the bone, to prevent stress risers with an appropriate working length. This procedure is unique, as it replaces the torn ligament with a natural substitute, in the appropriate location, through a minimally invasive procedure. This technique would be suitable for treatment of patients with either grade III or V acute acromioclavicular dislocations. Clinical outcomes for the first 13 consecutive patients treated with this procedure are reported, revealing excellent satisfaction rates with a Constant score of 96.6 at final follow-up.


Identification, characterization and expression profiling of cation-proton antiporter superfamily in Triticum aestivum L. and functional analysis of TaNHX4-B.
The monovalent cation proton antiporter (CPA) superfamily comprises Na+/H+ exchanger (NHX), K+ efflux antiporter (KEA), and cation/H+ exchanger (CHX) family proteins, which play vital functions in plants. A total of 107 TaCPA proteins were identified in Triticum aestivum, and phylogenetically classified into 35 TaNHX, 24 TaKEA and 48 TaCHX proteins. These families had representatives derived from all three sub-genomes. TaKEA genes consisted of higher number of exons, followed by TaNHXs and TaCHXs. The occurrence of about 10 transmembrane regions and higher composition of helices and coils support their membrane-bound and hydrophobic nature. Diverse expression in various tissues and modulated expression under stress conditions suggested their role in development and in response to stress. Co-expression analyses revealed their complex interaction networks. Expression of TaNHX4-B.1 and TaNHX4-B.4 facilitated differential abiotic stress tolerance to Escherichia coli. Our study provides comprehensive information about CPA genes, which would be useful in their future functional characterization.


Antibiotic and mammal IgG based lateral flow assay for simple and sensitive detection of Staphylococcus aureus.
Lateral flow assay (LFA), performed with simple devices and short detection time, is popular in field applications. Herein, a novel sandwich type-based LFA was constructed for high sensitivity and selectivity detection of Staphylococcus aureus (S. aureus). Vancomycin-immobilized gold nanoparticles (VAN-Au NPs) were utilized as the first identifier to capture S. aureus and the specificity was guaranteed by the second recognition agent of pig immunoglobulin G (IgG). In addition, gold growth was adopted for signal amplification to further improve the detection sensitivity. S. aureus could be directly assayed by this LFA within the concentration range of 1.0 × 103-1.0 × 108 cfu mL-1 with a detection limit of 1.0 × 103 cfu mL-1. Furthermore, the novel sandwich LFA realized S. aureus detection in food samples with admissible recoveries and established a rapid, simple, cost-effective and sensitive platform, could meet the demand for on-site testing of S. aureus.


Ambulatory blood pressure and blood lipids in a multiethnic sample of healthy adults.
Elevated blood pressure (BP), elevated serum cholesterol, and aberrant lipoprotein fractions (low levels of high-density lipoprotein (HDL) and high levels of low-density lipoprotein fractions and triglycerides) have all been used as measures that assess the "metabolic syndrome" and more recently in indexes of allostatic load, which are designed to assess the degree of integrated metabolic pathology. While there are ample data regarding the interrelationships of these measures in various pathophysiological settings, there are limited data regarding the interrelationship of ambulatory BP (ABP) and blood lipids in healthy subjects. The present study evaluates ABP-blood lipid relationships in a multiethnic sample of healthy adults. The subjects were 37 men (age = 40.9 ± 10.7 years) and 42 women (age = 35.8 ± 10.4 years) who were employed as hotel workers in Hawaii. Each wore an ABP monitor for one midweek workday and had pressures averaged in three daily microenvironments (work, home, and during sleep). They also had fasting blood samples taken for lipid profiling. Multivariate analysis of covariance shows that there was a strong inverse relationship between HDL and both systolic (P < 0.006) and diastolic (P < 0.006) BP, overall and in each microenvironment, but no statistically significant relationships with other lipid measures. These results suggest lipids and BP do not act as a group in healthy adults but that higher HDL is associated with lower BP. This latter finding is consistent with research that shows that HDL promotes vasodilation via its effect on endothelial nitric oxide synthase.


Structure of a polysaccharide from Providencia rustigianii O11 containing a novel amide of 2-acetamido-2-deoxygalacturonic acid with L-glutamyl-L-alanine.
An acidic polysaccharide was isolated from Providencia rustigianii O11 by the phenol-water extraction. The polysaccharide was cleaved by solvolysis with triflic acid to yield disaccharides with uronic acid derivatives at the non-reducing end. The polysaccharide and the disaccharides were studied by chemical analyses, high-resolution ESI MS, and 2D (1)H and (13)C NMR spectroscopy, and the following structure of the tetrasaccharide repeating unit of the polysaccharide was established: where GalNAcA stands for 2-acetamido-2-deoxygalacturonic acid, GalNAcA6GluAla for N-(2-acetamido-2-deoxygalacturonoyl)-l-glutam-1-yl-l-alanine, QuiNAc4NAcyl for 2-acetamido-4-[(S)-3-hydroxybutanoylamino]-2,4,6-trideoxyglucose (∼75%) or 2,4-diacetamido-2,4,6-trideoxyglucose (∼25%); the d configuration of GalNA and QuiN4N was ascribed tentatively. To the best of our knowledge, this is for the first time that an amide of uronic acid with a dipeptide is found in bacterial polysaccharides.


Phosphatidyl myo-inositol mannosides mimics built on an acyclic or heterocyclic core: synthesis and anti-inflammatory properties.
Phosphatidyl myo-inositol mannosides (PIMs) are constituents of the mycobacterial cell wall and possess immunomodulatory activities. Certain PIM derivatives have immunoprotective activity and are of interest as anti-inflammatory agents. In order to identify simplified analogues of PIMs that retain this interesting activity, we have prepared a series of new analogues based either on an acyclic or on a heterocyclic scaffold that replaces the inositol moiety, and evaluated these compounds for their inhibition of LPS-induced release of NO and pro-inflammatory cytokines by macrophages. It was found that the inositol moiety can be favourably replaced by an aza-cyclitol (trihydroxy-piperidine) or an oxa-cyclitol (trihydroxy-tetrahydropyran) unit, and that the configuration of the OH-carrying carbons does not play a significant role. The biological activity is reduced if the nitrogen atom is free in the aza-cyclitol unit.


Stakeholder expectations, experiences, and perceived outcomes: Implementation of a practice-integrated hospital pharmacist foundation residency program.
In 2017, a practice-integrated hospital pharmacist foundation residency program, modelling well-established international programs, was made available to all eligible hosting hospital pharmacy service providers in Australia. The South Australian public hospital pharmacy service implemented the nation's first accredited statewide residency model that also incorporates enrolment and completion of a tailored post-graduate qualification. This study aimed to explore stakeholder perspectives pertaining to implementation and adoption of the South Australian public hospital pharmacy statewide residency program. Eligible participants were recruited via purposeful sampling. Stakeholders overseeing preceptors and residents as well as those with authority to influence decision-making for future program planning were invited to participate in semi-structured interviews. Each interview was audio-recorded, transcribed, and organized using qualitative data analysis software. Reflexive thematic analysis was applied to data synthesis, and findings were discussed through the lens of the PRECEDE logic framework. Thirty-three participants were consented and de-identified. Three themes were identified: resource capacity for program sustainability; current and expected future impact on workforce development and career progression; and conflicts around uniformity, selectivity, and individual motivations. Research findings affirmed program benefits to overall workforce development whilst highlighting persisting program sustainability dilemmas. Findings have highlighted the need for a cost benefit analysis of each aspect of the residency program and the importance of tailoring to individual and site differences. Several strategies were recommended to optimize resources for increased capacity, to better support residents and preceptors, and to reduce program intensity.


A phase II study of cisplatin therapy in recurrent childhood brain tumors. A report from the Childrens Cancer Study Group.
Thirty-six children with brain tumors were treated with surgery, radiation and/or adjuvant chemotherapy. After tumor recurrence, cisplatin (60 mg/m2/day IV X 2) was given every three to four weeks. CT scans were used to measure drug response prior to the first, third and fifth courses. Complete and partial responses were demonstrated in nine of 31 evaluable patients. Dose limiting toxicities were renal and auditory. Seven patients developed the syndrome of inappropriate antidiuretic hormone secretion. This study confirms that cisplatin is active in a spectrum of brain tumors.


Systemic administration of the potent mGlu8 receptor agonist (S)-3,4-DCPG induces c-Fos in stress-related brain regions in wild-type, but not mGlu8 receptor knockout mice.
The effect of a novel and potent metabotropic glutamate 8 (mGlu8) receptor agonist, (S)-3,4-dicarboxyphenylglycine (DCPG), was studied in vivo in mouse brain. c-Fos expression was used as a marker of neuronal activity in specific brain regions 2 h after systemic (S)-3,4-DCPG (3-100 mg/kg, i.p.). The selectivity of (S)-3,4-DCPG on mGlu8 receptors was determined in mGlu8 receptor knockout mice. In wild-type mice, (S)-3,4-DCPG (100 mg/kg) significantly increased c-Fos expression in several stress-related brain regions: paraventricular nucleus of the hypothalamus, central nucleus of the amygdala, lateral parabrachial nucleus and locus coeruleus. In the central nucleus of the amgydala, more than 92% of c-Fos positive neurons were identified as GABAergic inhibitory neurons after (S)-3,4-DCPG. Moreover, (S)-3,4-DCPG significantly induced c-Fos in the superficial gray layer of the superior colliculus, a central visual region. c-Fos expression was unchanged by (S)-3,4-DCPG in mGlu8 receptor knockout mice. Our results indicate that systemic (S)-3,4-DCPG alters neuronal excitability in specific brain regions via mGlu8 receptor. The prominent activation of stress areas suggests a role for mGlu8 receptors in the central integration of stress responses. Furthermore, our results indicate that systemic (S)-3,4-DCPG can be used as a tool to explore behavioral and cellular consequences of mGlu8 receptor activation.


Clinical presentation of intracranial metastases.
Intracranial metastases are the single most costly neurologic ailment. This article provides the clinician with detailed statements of the patterns of their appearance and tumors of origin. Most common causative cancers are of lung, breast, or melanoma origin. Aggressive therapies carry with them the risk of nervous system dissemination of rare tumors, remote complications of these cancers and the morbidity of therapy.


Repetitive genomic elements in Campomanesia xanthocarpa: prospection, characterization and cross amplification of molecular markers.
Repetitive genomic elements were prospected in Campomanesia xanthocarpa, aiming to characterize these elements in a non-model plant species and to develop species-specific microsatellite markers. Approximately 4.12% of the partial genome of C. xanthocarpa is composed of repetitive elements, being retrotransposons the most widely represented. A total of nine polymorphic microsatellite markers were obtained: four nuclear-neutral, two nuclear EST, two plastidial and one mitochondrial. Levels of population genetic diversity of four natural populations of C. xanthocarpa were characterized using these markers. In addition, the cross-species amplification of the microsatellite markers was tested in seven species of tribe Myrteae (Myrtaceae). The characterized microsatellite markers revealed low to moderate levels of genetic diversity (expected heterozygosity range: 0.33-0.57; observed heterozygosity: 0.26-0.74 and number of alleles: 2.25-4.25). Cross-species amplification was successful for all loci, except Cxant76. These nine markers will contribute for studies on genetic diversity, gene flow, plant selection and breeding of this species, towards the conservation of natural populations, as well as its commercial use.


Transfected human neuropeptide Y cDNA expression in mouse pituitary cells. Inducible high expression, peptide characterization, and secretion.
An expression vector was constructed that placed the cDNA for human neuropeptide Y (NPY) under the control of the mouse metallothionein promoter and was used to transfect the AtT-20 mouse anterior pituitary corticotrope cell line. AtT-20 cells normally process the pro-ACTH/endorphin precursor but do not produce detectable levels of NPY. The resulting AtT-20/NPY cell line (Mt.NPY1a) was used to study the ability of the corticotrope cells to synthesize, process, and secrete the foreign proNPY-related peptide products. The stable cell line created contains approximately 40 copies of proNPY cDNA per cell. NPY mRNA levels and proNPY synthesis were increased at least 35-fold when maximally induced with cadmium; proNPY synthesis was also induced by glucocorticoids. Upon induction the NPY secretion rate was equimolar to that of the endogenous peptides. ProNPY, NPY, and the COOH-terminal peptide produced by this cell line had molecular weight and amino acid-labeling pattern predicted from cDNA sequence data and from previous isolation of NPY-related molecules from NPY-producing cells. The structures of secreted proNPY, NPY, and COOH-terminal peptide, as well as determination of the site of proteolytic cleavage between NPY and the COOH-terminal peptide, were determined by tryptic mapping and Edman degradation of secreted biosynthetically labeled peptide products. The proNPY molecule appears to be processed in the same pathway responsible for cleavage of the endogenous pro-ACTH/endorphin precursor. Secretion of proNPY-derived peptides paralleled secretion of endogenous pro-ACTH/endorphin-derived products, under both basal and stimulated conditions. With induction proNPY expression there is a dose-dependent inhibition of both proNPY and pro-ACTH/endorphin proteolytic processing.


Distribution patterns of stromal eosinophil cells in chick thymus during postnatal development.
Eosinophils are a type of thymic stromal cell that are present in the thymus of both humans and mice. They participate in regulating T-cell development under non-pathological conditions. However, studies are scarce regarding the role of eosinophils in the development of the thymus in chickens. Therefore, this study investigated the distribution of eosinophils in normal chicken thymi at different stages of development. Seven thymi were obtained from chickens at days 1, 21 and 35 of development. The distribution of eosinophils in the thymi was analyzed by histological and immunohistochemical techniques using Lendrum's chromotrope 2R method and an antibody against eosinophilic cationic protein (ECP), respectively. Eosinophils were constitutively located in the chick thymus. They were mainly distributed in the thymic corticomedullary junction and medulla, especially around vessels and Hassall's corpuscles, and only a few were in the trabeculae among thymic lobules and around vessels. There were none in the cortex. The number of thymic eosinophils decreased with increasing age (P<0.01). These results indicated that eosinophils comprise a type of thymic stromal cells in the chick, which may regulate thymic development, especially during the early stages of development.


Formation by phospholipase A2 of prostaglandins and thromboxane A2-like activity in the platelets of normal and essential fatty acid deficient rats. Comparison with effect on human and rabbit platelets.
When rat platelets are incubated with phospholipase A2, thromboxane A2-like activity and prostaglandins are formed. The amounts are approximately similar, whether aggregation is induced after the incubation or not. No aggregation is observed when the platelets are incubated with phospholipase A2. In the platelets of essential fatty acid deficient rats, only small amounts of thromboxane A2-like acitivity and prostaglandins are formed. No formation of these substances occurs when human and rabbit platelets are incubated with phospholipase A2. The results indicate that formation of thromboxane A2-like activity enhances aggregation in rat platelets, but that aggregation is not induced.


In vivo bone and cartilage MRI using fully-balanced steady-state free-precession at 7 tesla.
The purpose of this work was to investigated the feasibility of fully-balanced steady-state free-precession (bSSFP) pulse sequence for trabecular bone and knee cartilage imaging in vivo using ultra-high-field (UHF) MRI at 7T in comparison with pulse sequences previously used at 3T. We showed that bSSFP and spin-echo imaging is possible at higher field strengths within 3.2 W/kg specific absorption rate (SAR) constraints. All pulse sequences were numerically optimized based on measured tissue relaxation parameters from six healthy volunteers (T(1) = 820 +/- 128 ms, T(2) = 43.5 +/- 3 ms for bone marrow and T(1) = 1745 +/- 104 ms and T(2) = 30 +/- 4 ms for cartilage). From simulations of the Bloch equation, a signal-to-noise ratio (SNR) increase of more than 1.9 was predicted. Cartilage SNR of bSSFP was 2.4 times higher at 7T (51.3 +/- 4.3) compared with 3T (21.3 +/- 3.3). Bone SNR increased from 11.8 +/- 2.0 to 13.2 +/- 2.5 at the higher field strength. We concluded that there is SNR benefit and great potential for bone and cartilage imaging at higher field strength.


Purine metabolism in Leishmania donovani amastigotes and promastigotes.
Purine metabolism in Leishmania donovani amastigotes was found to be similar to that of promastigotes with the exception of adenosine metabolism. Adenosine kinase activity in amastigotes is approximately 50-fold greater than in promastigotes. Amastigotes deaminate adenosine to inosine through adenosine deaminase, an enzyme not present in promastigotes. Inosine is cleaved to hypoxanthine and phosphoribosylated by hypoxanthine-guanine phosphoribosyltransferase. Promastigotes cleave adenosine to adenine and deaminate adenine to hypoxanthine via adenase, an enzyme not present in amastigotes. Hypoxanthine is phosphoribosylated by hypoxanthine-guanine phosphoribosyltransferase.


Synthesis, Aromaticity, and Application of peri-Pentacenopentacene: Localized Representation of Benzenoid Aromatic Compounds.
We report the synthesis and optoelectronic properties of TIPS-peri-pentacenopentacene (TIPS-PPP), a vertical extension of TIPS-pentacene (TIPS-PEN) and a low-band-gap material with remarkable stability. We found the synthetic conditions to avoid the competition between 1,2- and 1,4-addition of lithium acetylide on the large aromatic dione. The high stability of TIPS-PPP is due to the peri-fusion which increases the aromaticity by generating two localized aromatic sextets that are flanked with 2 diene fragments, similar to two fused-anthracenes. Like TIPS-PEN, TIPS-PPP shows the archetypal 2D brickwall motif in crystals with a larger transfer integral and smaller reorganization energy. The high mobility of up to 1 cm2 V-1 s-1 was obtained in an organic field-effect transistor fabricated by a wet process. Also, TIPS-PPP was used as a near-infrared (NIR) emitter for NIR organic-light-emitting-diode devices resulting in a high external quantum efficiency at 800 nm.


Aberrant forebrain signaling during early development underlies the generation of holoprosencephaly and coloboma.
In this review, we highlight recent literature concerning the signaling mechanisms underlying the development of two neural birth defects, holoprosencephaly and coloboma. Holoprosencephaly, the most common forebrain defect, occurs when the cerebral hemispheres fail to separate and is typically associated with mispatterning of embryonic midline tissue. Coloboma results when the choroid fissure in the eye fails to close. It is clear that Sonic hedgehog (Shh) signaling regulates both forebrain and eye development, with defects in Shh, or components of the Shh signaling cascade leading to the generation of both birth defects. In addition, other intercellular signaling pathways are known factors in the incidence of holoprosencephaly and coloboma. This review will outline recent advances in our understanding of forebrain and eye embryonic pattern formation, with a focus on zebrafish studies of Shh and retinoic acid pathways. Given the clear overlap in the mechanisms that generate both diseases, we propose that holoprosencephaly and coloboma can represent mild and severe aspects of single phenotypic spectrum resulting from aberrant forebrain development. This article is part of a Special Issue entitled Zebrafish Models of Neurological Diseases.


Real-world effectiveness of intensive chemotherapy with 7&3 versus venetoclax and hypomethylating agent in acute myeloid leukemia.
Intensive chemotherapy with cytarabine and anthracycline (7&3) remains the standard therapy for patients medically fit for induction, but the assessment of fitness remains controversial. Venetoclax and hypomethylating agent (ven/HMA) combination therapy has improved outcomes in unfit patients but no prospective study has assessed ven/HMA versus 7&3 as initial therapy in older, fit patients. Given no studies and expectation of ven/HMA use in patients outside of trial criteria, we evaluated retrospective outcomes among newly diagnosed patients. A nationwide electronic health record (EHR)-derived database and the University of Pennsylvania EHR identified 312 patients receiving 7&3 and 488 receiving ven/HMA who were 60-75 years old without history of organ failure. Ven/HMA patients were older and more likely to have secondary AML, adverse cytogenetics, and adverse mutations. Median overall survival (OS) for patients receiving intensive chemotherapy was 22 versus 10 months for ven/HMA (HR 0.53, 95% CI 0.40-0.60). Controlling for measured baseline characteristic imbalances reduced survival advantage by half (HR 0.71, 95% CI 0.53-0.94). A sub-group of patients with equipoise, likelihood at least 30%-70% of receiving either treatment, had similar OS outcomes (HR 1.10, 95% CI 0.75-1.6). Regarding safety outcomes, 60-day mortality was higher for ven/HMA (15% vs. 6% at 60 days) despite higher documented infections and febrile neutropenia for 7&3. In this multicenter real-word dataset, patients selected for intensive chemotherapy had superior OS but a large group had similar outcomes with ven/HMA. Prospective randomized studies, controlling for both measured and unmeasured confounders, must confirm this outcome.


The promise of PROMIS(®) for addiction.
The field of addiction treatment has a measurement problem that pervades efforts to help patients achieve self-sustainable recovery. The impact of using older measurement technology has increased the measurement burden on both service providers and patients, while at the same time limiting the scope and frequency of measurement. The resulting burden can affect provider performance, patient access, and addiction recovery. This paper underscores the need for applying modern measurement theory techniques to reduce the measurement burden currently affecting most if not all major aspects of treatment and recovery. It is currently possible to obtain information more precisely, over a broad spectrum of recovery-oriented domains, faster and at lower cost than current measurement practices allow. However, a persistent research effort will be necessary to achieve that goal.


3D Decellularized Native Extracellular Matrix Scaffold for In Vitro Culture Expansion of Human Wharton's Jelly-Derived Mesenchymal Stem Cells (hWJ MSCs).
Mesenchymal stem cells (MSCs) are derived from Wharton's jelly tissue of the human umbilical cord. Given appropriate culture conditions, these cells can self-renew and differentiate into multiple cell types across the lineages. Among the properties exhibited by these cells, immunomodulation through secretion of trophic factors has been widely exploited in a broad spectrum of preclinical/clinical regenerative applications. Moreover, the extracellular matrix is found to play a major role apart from niche cells in determining stem cell fate including that of MSCs. Therefore, the currently employed technique of two-dimensional culture expansion can alter the inherent properties of naïve MSCs originally residing within the three-dimensional space. This limitation can be overcome to some extent by using native extracellular matrix scaffold culture system which mimics the in situ microenvironment. In this chapter, we have elucidated the protocol for the preparation of a native extracellular matrix scaffold by decellularization of the MSC sheet and thereof culture expansion and characterization of human Wharton's jelly-derived MSCs.