A phagocytosis-enhancing factor in human plasma.
A phagocytosis-enhancing factor (PEF) with the capacity to stimulate the ingestion of sensitized sheep erythrocytes by human polymorphonuclear and mononuclear leucocytes has been isolated from human plasma by chromatography on DEAE-cellulose and filtration on Sephadex G-150 and Sephadex G-100. PEF is a protein of approximately 70,000 molecular weight which is susceptible to inactivation by heating at 60 degrees or by tryptic digestion. PEF promotes phagocytosis of erythrocytes sensitized with intact 7S antibody or bearing the C3b complement fragment, but not of unsensitized erythrocytes or erythrocytes sensitized with 19S antibody. The specificity of PEF interaction with target erythrocytes and the persistence of its stimulatory effect after the target cells are washed suggest that it promotes phagocytosis by an action on the erythrocytes.


Improved Documentation of Electronic Cigarette Use in an Electronic Health Record.
The use of electronic cigarettes (e-cigarettes) can affect patient health and clinical care. However, the current documentation of e-cigarette use in the electronic health records (EHR) is inconsistent. This report outlines how the ambulatory clinical practices of a large U.S. hospital system optimized its electronic health records (EHR) framework to better record e-cigarettes used by patients. The new EHR section for e-cigarette information was implemented for outpatient appointments. During a 30-week evaluation period post-implementation, 638,804 patients (12 yrs and older) completed ambulatory appointments within the health system; of these, the new section contained e-cigarette use information for 37,906 (6%) patients. Among these patients, 1005 (2.7%) were identified as current e-cigarette users (current every day or current some day e-cigarette use), 941 (2.5%) were reported as former e-cigarette users, and 35,960 (94%) had never used e-cigarettes. A separate EHR section to document e-cigarette use is feasible within existing clinical practice models. Utilization of the new section was modest in routine clinical practice, indicating the need for more intensive implementation strategies that emphasize the health effects of e-cigarette use, and how consistent ascertainment could improve clinical practice.


[Mitokorrektine stimulates angiogenesis in vitro].
The effect of mitokorrectine (complex native oligopeptides isolated from neonatal pig brain) on endothelial cells in culture was investigated. It was revealed that the drug concentration-dependently induces angiogenesis in vitro. Mitogen effect of Mitokorrectine was shown by MTT-test and routine cell count in concentration diapason (0.1-1 mg/ml) which means an increased number of cells by 25 +/- 5% and cell subpopulation of proliferative pool (G2/M+S) 1,8 times in concentration diapason mitokorrectine (0.01-0.05 mg/ml) in comparison with control. In 3-D culture and in stationary phase we detected induction of differentiation of endothelial cells, a decrease the level of NO production and enhancement of glucose metabolism and stimulation of formation of capillary-like tubes.


Perceptual and Cognitive Factors Imposing "Speed Limits" on Reading Rate: A Study with the Rapid Serial Visual Presentation.
Adults read at high speed, but estimates of their reading rate vary greatly, i.e., from 100 to 1500 words per minute (wpm). This discrepancy is likely due to different recording methods and to the different perceptual and cognitive processes involved in specific test conditions. The present study investigated the origins of these notable differences in RSVP reading rate (RR). In six experiments we investigated the role of many different perceptual and cognitive variables. The presence of a mask caused a steep decline in reading rate, with an estimated masking cost of about 200 wpm. When the decoding process was isolated, RR approached values of 1200 wpm. When the number of stimuli exceeded the short-term memory span, RR decreased to 800 wpm. The semantic context contributed to reading speed only by a factor of 1.4. Finally, eye movements imposed an upper limit on RR (around 300 wpm). Overall, data indicate a speed limit of 300 wpm, which corresponds to the time needed for eye movement execution, i.e., the most time consuming mechanism. Results reconcile differences in reading rates reported by different laboratories and thus provide suggestions for targeting different components of reading rate.


Lower extremity joint kinetic responses to external resistance variations.
The purpose of this investigation was to determine if increases in external resistance during a squat movement would be controlled by proportionally scaling the net joint moment work or average net joint moment (NJM) at the hip, knee, and ankle. Eighteen experienced subjects performed 3 sets of 3 repetitions each of a squat movement using resistances of 25, 50, 75, and 100% of their 3-repetition maximum, while instrumented for biomechanical analyses. Standard inverse dynamics techniques and numerical integration were used to calculate the NJM work and average NJM of each joint. A combination of single-subject and group mean statistical analyses indicated that the neither the NJM work nor average NJM increased proportionately in response to increases in external loading. Results suggest a complex control strategy in which the hip was the dominant contributor, increased linearly with the external load, and had low variability. The knee and ankle contributions were neither as great nor as linear, and were highly variable, suggesting that they were influenced by more than just the external load. The disproportionate response of each joint to varying external resistances suggests that controlling the force output of multijoint chains requires further study and modifications to existing motor control theories.


Purification and properties of alpha-mannosidase from bakers' yeast.
The yeast alpha-mannosidase [EC 220.127.116.11] was purified 1160-fold from the crude extract of the autolysate. The purified preparation was practically free from alpha-glucosidase, beta-glucosidase, alpha-galactosidase, beta-galactosidase, beta-mannosidase, and beta-N-acetylhexosaminidase activities. After the separation of yeast mannan during the purification procedures the enzyme became unstable but could be stored at 5 degrees C for three weeks with 50% loss of activity. The purified enzyme hydrolyzed both aryl and alkyl mannosides, but hydrolysis of yeast mannan proceeded slowly. Yeast mannan and Zn2+ increased the enzyme catalyzed hydrolysis of p-nitrophenyl mannoside, whereas NaN3, monoiodoacetate and methyl alpha-D-mannoside acted as inhibitors. The molecular weight was estimated to be 450,000 by gel filtration.


Changes in the titre of serum opsonins and phagocytic properties of mouse peritoneal macrophages following injection of endotoxin.
A study has been made of the changes in the opsonic properties of mouse serum and bactericidal activity of mouse peritoneal macrophages at different times following the injection of various doses of lipopolysaccharide. It has been found that changes in the percentage of bacteria phagocytosed and killed within the mouse peritoneal cavity in a given time could be correlated with changes in the opsonic titre of the serum. Macrophages harvested from the peritoneal cavities of mice injected with endotoxin appear to be more efficient in phagocytosing bacteria in the presence of serum opsonins than macrophages obtained from normal mice. The relative importance of these changes in determining an animal's resistance to infection is discussed.


An allosteric inhibitor against the therapy-resistant mutant forms of EGFR in non-small cell lung cancer.
Epidermal growth factor receptor (EGFR) therapy using small-molecule tyrosine kinase inhibitors (TKIs) is initially efficacious in patients with EGFR-mutant lung cancer, although drug resistance eventually develops. Allosteric EGFR inhibitors, which bind to a different EGFR site than existing ATP-competitive EGFR TKIs, have been developed as a strategy to overcome therapy-resistant EGFR mutations. Here we identify and characterize JBJ-09-063, a mutant-selective allosteric EGFR inhibitor that is effective across EGFR TKI-sensitive and resistant models, including those with EGFR T790M and C797S mutations. We further uncover that EGFR homo- or heterodimerization with other ERBB family members, as well as the EGFR L747S mutation, confers resistance to JBJ-09-063, but not to ATP-competitive EGFR TKIs. Overall, our studies highlight the potential clinical utility of JBJ-09-063 as a single agent or in combination with EGFR TKIs to define more effective strategies to treat EGFR-mutant lung cancer.


Activating PIK3CA mutations coexist with BRAF or NRAS mutations in a limited fraction of melanomas.
Activated PI3K-AKT pathway may contribute to decrease sensitivity to inhibitors of key pathogenetic effectors (mutated BRAF, active NRAS or MEK) in melanoma. Functional alterations are deeply involved in PI3K-AKT activation, with a minimal role reported for mutations in PIK3CA, the catalytic subunit of the PI3K gene. We here assessed the prevalence of the coexistence of BRAF/NRAS and PIK3CA mutations in a series of melanoma samples. A total of 245 tumor specimens (212 primary melanomas and 33 melanoma cell lines) was screened for mutations in BRAF, NRAS, and PIK3CA genes by automated direct sequencing. Overall, 110 (44.9%) samples carried mutations in BRAF, 26 (10.6%) in NRAS, and 24 (9.8%) in PIK3CA. All identified PIK3CA mutations have been reported to induce PI3K activation; those detected in cultured melanomas were investigated for their interference with the antiproliferative activity of the BRAF-mutant inhibitor vemurafenib. A reduced suppression in cell growth was observed in treated cells carrying both BRAF and PIK3CA mutations as compared with those presenting a mutated BRAF only. Among the analysed melanomas, 12/245 (4.9%) samples presented the coexistence of PIK3CA and BRAF/NRAS mutations. Our study further suggests that PIK3CA mutations account for a small fraction of PI3K pathway activation and have a limited impact in interfering with the BRAF/NRAS-driven growth in melanoma.


Coexistence of two functioning T-cell repertoires in healthy ex-thalassemics bearing a persistent mixed chimerism years after bone marrow transplantation.
Bone marrow transplantation (BMT) from an HLA-identical donor is an established therapy to cure homozygous beta-thalassemia. Approximately 10% of thalassemic patients developed a persistent mixed chimerism (PMC) after BMT characterized by stable coexistence of host and donor cells in all hematopoietic compartments. Interestingly, in the erythrocytic lineage, close to normal levels of hemoglobin can be observed in the absence of complete donor engraftment. In the lymphocytic lineage, the striking feature is the coexistence of immune cells. This implies a state of tolerance or anergy, raising the issue of immunocompetence of the host. To understand the state of the T cells in PMC, repertoire analysis and functional studies were performed on cells from 3 ex-thalassemics. Repertoire analysis showed a profound skewing. This was due to an expansion of some T cells and not to a collapse of the repertoire, because phytohemagglutinin stimulation showed the presence of a complex repertoire. The immunocompetence of the chimeric immune systems was further established by showing responses to alloantigens and recall antigens in vitro. Both host and donor lymphocytes were observed in the cultures. These data suggest that the expanded T cells play a role in specific tolerance while allowing a normal immune status in these patients.


[Parameters of heart rhythm variability and QT-interval in patients with ischemic heart disease and type 2 diabetes].
We studied parameters of heart rhythm variability and QT intervals in 141 patients (mean age 52.4 +/- 7.3 years) with unstable angina and acute myocardial infarction; 101 of these patients (71.6%) had type 2 diabetes. We found that in patients with diabetes parameters RRNN, LF and LF/HF were significantly lower. In patients with diabetes and risk of microangiopathy lowering of LF was maximal and correlated with elevation of glycosylated hemoglobin. In patients with diabetes and neuropathy significant reduction of power in HF range and increase of maximal corrected QT-interval value were observed. It is possible, that in patients with ischemic heart disease type 2 diabetes causes decrease of tone of vegetative nervous system mainly due to activity of parasympathetic part and this is accompanied by impaired repolarization of cardiac ventricles.


Production of resveratrol in recombinant microorganisms.
Resveratrol production in Saccharomyces cerevisiae was compared to that in Escherichia coli. In both systems, 4-coumarate:coenzyme A ligase from tobacco and stilbene synthase from grapes were expressed. When p-coumaric acid was used as the precursor, resveratrol accumulations in the culture medium were observed to be comparable in E. coli (16 mg/liter) and yeast (6 mg/liter).


Confounding mitigation for the exposure-response relationship of bevacizumab in colorectal cancer patients.
The exposure-response relationship of bevacizumab may be confounded by various factors, including baseline characteristics, time-dependent target engagement and recursive relationships between exposure and response, requiring effective mitigation. This study aimed at investigating the exposure-response relationships of bevacizumab in mCRC patients while mitigating potential biases. Bevacizumab pharmacokinetics was described using target-mediated drug disposition (TMDD) modeling. Relationships between target kinetics, and progression-free (PFS) and overall (OS) survivals were assessed using joint pharmacokinetic and parametric hazard function models. Both potential biases due to prognostic-driven and response-driven of the concentration-effect relationship were mitigated. These models evaluated the impact of increased antigen target levels,clearance, and intensified dosing regimen on survival. Estimated target-mediated pharmacokinetic parameters in 130 assessed patients were baseline target levels (R0 =8.4 nM), steady-state dissociation constant (KSS =10 nM) and antibody-target complexes elimination constant (kint =0.52 day-1 ). The distribution of R0 was significantly associated with increased baseline concentrations of carcinoembryonic antigen, circulating Vascular Endothelial Growth Factor, and the presence of extra-hepatic metastases. Unbound target levels (R) significantly influenced both progression and death hazard functions. Increasing baseline target levels and/or clearance values led to decreased bevacizumab unbound concentrations, increased R levels, and shortened PFS and OS, while increasing bevacizumab dose led to decreased R and longer survival. This study is the first to demonstrate the relationship between bevacizumab concentrations, target involvement and clinical efficacy by effectively mitigating potential sources of bias. Most of target amount may be tumoral in mCRC. Future studies should provide a more in-depth description of this relationship.


Insights into Biofilm Dispersal Regulation from the Crystal Structure of the PAS-GGDEF-EAL Region of RbdA from Pseudomonas aeruginosa.
RbdA is a positive regulator of biofilm dispersal of Pseudomonas aeruginosa Its cytoplasmic region (cRbdA) comprises an N-terminal Per-ARNT-Sim (PAS) domain followed by a diguanylate cyclase (GGDEF) domain and an EAL domain, whose phosphodiesterase activity is allosterically stimulated by GTP binding to the GGDEF domain. We report crystal structures of cRbdA and of two binary complexes: one with GTP/Mg2+ bound to the GGDEF active site and one with the EAL domain bound to the c-di-GMP substrate. These structures unveil a 2-fold symmetric dimer stabilized by a closely packed N-terminal PAS domain and a noncanonical EAL dimer. The autoinhibitory switch is formed by an α-helix (S-helix) immediately N-terminal to the GGDEF domain that interacts with the EAL dimerization helix (α6-E) of the other EAL monomer and maintains the protein in a locked conformation. We propose that local conformational changes in cRbdA upon GTP binding lead to a structure with the PAS domain and S-helix shifted away from the GGDEF-EAL domains, as suggested by small-angle X-ray scattering (SAXS) experiments. Domain reorientation should be facilitated by the presence of an α-helical lever (H-helix) that tethers the GGDEF and EAL regions, allowing the EAL domain to rearrange into an active dimeric conformation.IMPORTANCE Biofilm formation by bacterial pathogens increases resistance to antibiotics. RbdA positively regulates biofilm dispersal of Pseudomonas aeruginosa The crystal structures of the cytoplasmic region of the RbdA protein presented here reveal that two evolutionarily conserved helices play an important role in regulating the activity of RbdA, with implications for other GGDEF-EAL dual domains that are abundant in the proteomes of several bacterial pathogens. Thus, this work may assist in the development of small molecules that promote bacterial biofilm dispersal.


Liver colour scoring index, carotenoids and lipid content assessment as a proxy for lumpfish (Cyclopterus lumpus L.) health and welfare condition.
Ensuring lumpfish health and welfare in salmon farms is vital to reduce the high mortality rates reported and to guarantee a high delousing efficiency. Recent observations of farmed lumpfish livers have shown colours ranging from pale (colours 1 and 2), through bright orange (colours 3 and 4), to dark reddish-brown (colours 5 and 6), some of which may be related to welfare condition. To characterize the status of lumpfish deployed in four Faroese salmon farms, several welfare indicators were assessed: a weight-length relationship, scoring of external physical damage, and after dissection, stomach content and liver colour scoring. Liver samples were weighed, stored and analysed for lipid content, lipid classes, total pigments, fatty acid profile and histopathology to explain the differences between the mentioned liver colours. Bright orange livers, liver colours 3 and 4, were related to increased levels of carotenoid pigments rather than levels of lipids and appear to reflect good fish welfare. However, dark reddish-brown colours, liver colours 5 and 6, were associated with very low levels of triacyl glycerides in the liver, indicating use of lipid reserves and poor welfare condition. Histopathology confirmed that the dark reddish-brown livers, liver colours 5 and 6, formed a distinct group. Thus, liver colour was shown to be a good welfare indicator and should be monitored in farms.


Substance P and cholecystokinin-like immunoreactive varicosities in somatosensory and autonomic regions of the rat spinal cord: a quantitative study of coexistence.
Substance P- and cholecystokinin-like immunoreactivities have been shown to coexist in some, but not all, dorsal root ganglion cell bodies of the rat. Quantitative immunofluorescence techniques were used in the present study to describe densities of substance P- and cholecystokinin-like immunoreactive varicosities in several spinal cord nuclei. By combining simultaneous immunofluorescent techniques on one tissue section with computerized image processing, coexistence of substance P- and cholecystokinin-like immunoreactivity in varicosities was also quantified. By comparing spinal cord regions between normal and unilateral dorsal rhizotomy affected animals, densities of substance P- and cholecystokinin-like immunoreactive varicosities associated with primary afferent neurons were established. To determine the densities of immunoreactive varicosities that were related to unmyelinated primary afferent fibers, data were compared between normal animals and those treated neonatally with capsaicin. Four major observations were made: (1) Substance P- and cholecystokinin-like immunoreactivity coexist in populations of varicosities in sensory and autonomic regions of rat spinal segment L6. (2) Within the superficial laminae of the dorsal horn, varicosities containing both substance P- and cholecystokinin-like immunoreactivity and cholecystokinin-like immunoreactivity alone are of primary afferent neuron origin, but those containing only substance P-like immunoreactivity are most likely of spinal or descending neuronal origin. (3) Capsaicin-insensitive cholecystokinin-like immunoreactive varicosities were present predominantly in lamina I. These data suggest some cholecystokinin-like immunoreactive varicosities are associated with myelinated primary afferent neurons. (4) Primary afferent fibers containing substance P- and cholecystokinin-like immunoreactivity project to intermediate gray regions of the rat spinal cord. A large proportion of these fibers are capsaicin sensitive, suggesting that they are unmyelinated.


Differences in the ability of human T-cell lymphotropic virus type 1 (HTLV-1) and HTLV-2 tax to inhibit p53 function.
We have analyzed the functional activity of the p53 tumor suppressor in human T-cell lymphotropic virus type 2 (HTLV-2)-transformed cells. Abundant levels of the p53 protein were detected in both HTLV-2A and -2B virus-infected cell lines. The p53 was functionally inactive, however, both in transient-transfection assays using a p53 reporter plasmid and in induction of p53-responsive genes in response to gamma irradiation. We further investigated HTLV-2A Tax and HTLV-2B Tax effects on p53 activity. Interestingly, although Tax-2A and -2B inactivate p53, the Tax-2A protein appears to inhibit p53 function less efficiently than either Tax-1 or Tax-2B. In transient-cotransfection assays, Tax-1 and Tax-2B inactivated p53 by 80%, while Tax2A reduced p53 activity by 20%. In addition, Tax-2A does not increase the steady-state level of cellular p53 as well as Tax-1 or -2B does in the same assays. Cotransfection assays demonstrated that Tax-2A could efficiently transactivate CREB-responsive promoters to the same level as Tax-1 and Tax-2B, indicating that the protein was functional. This report provides evidence of the first functional difference between the HTLV-2A and -2B subtypes. This comparison of the action of HTLV-1 and HTLV-2 Tax proteins on p53 function will provide important insights into the mechanism of HTLV transformation.


Radiation-induced versus endogenous DNA damage: possible effect of inducible protective responses in mitigating endogenous damage.
Ionizing radiation (IR) causes damage to DNA that is apparently proportional to absorbed dose. The incidence of radiation-induced cancer in humans unequivocally rises with the value of absorbed doses above about 300 mGy, in a seemingly linear fashion. Extrapolation of this linear correlation down to zero-dose constitutes the linear-no-threshold (LNT) hypothesis of radiation-induced cancer incidence. The corresponding dose-risk correlation, however, is questionable at doses lower than 300 mGy. Non-radiation induced DNA damage and, in consequence, oncogenic transformation in non-irradiated cells arises from a variety of sources, mainly from weak endogenous carcinogens such as reactive oxygen species (ROS) as well as from micronutrient deficiencies and environmental toxins. In order to relate the low probability of radiation-induced cancer to the relatively high incidence of non-radiation carcinogenesis, especially at low-dose irradiation, the quantitative and qualitative differences between the DNA damages from non-radiation and radiation sources need to be addressed and put into context of physiological mechanisms of cellular protection. This paper summarizes a co-operative approach by the authors to answer the questions on the quantitative and qualitative DNA damages from non-radiation sources, largely endogenous ROS, and following exposure to low doses of IR. The analysis relies on published data and justified assumptions and considers the physiological capacity of mammalian cells to protect themselves constantly by preventing and repairing DNA damage. Furthermore, damaged cells are susceptible to removal by apoptosis or the immune system. The results suggest that the various forms of non-radiation DNA damage in tissues far outweigh corresponding DNA damage from low-dose radiation exposure at the level of, and well above, background radiation. These data are examined within the context of low-dose radiation induction of cellular signaling that may stimulate cellular protection systems over hours to weeks against accumulation of DNA damage. The particular focus is the hypothesis that these enhanced and persisting protective responses reduce the steady state level of non-radiation DNA damage, thereby reducing deleterious outcomes such as cancer and aging. The emerging model urgently needs rigorous experimental testing, since it suggests, importantly, that the LNT hypothesis is invalid for complex adaptive systems such as mammalian organisms.


Inositol-requiring mutants of Saccharomyces cerevisiae.
Fifty-two inositol-requiring mutants of Saccharomyces cerevisiae were isolated following mutagenesis with ethyl methanesulfonate. Complementation and tetrad analysis revealed ten major complementation classes, representing ten independently segregating loci (designated ino1 through ino10) which recombined freely with their respective centromeres. Members of any given complementation class segregated as alleles of a single locus. Thirteen complementation subclasses were identified among thirty-six mutants which behaved as alleles of the ino1 locus. The complementation map for these mutants was circular. - Dramatic cell viability losses indicative of unbalanced growth were observed in liquid cultures of representative mutants under conditions of inositol starvation. Investigation of the timing, kinetics, and extent of cell death revealed that losses in cell viability in the range of 2-4 log orders could be prevented by the addition of inositol to the medium or by disruption of protein synthesis with cycloheximide. Mutants defective in nine of the ten loci identified in this study displayed these unusual characteristics. The results suggest an important physiological role for inositol that may be related to its cellular localization and function in membrane phospholipids. The possibility is discussed that inositol deficiency initiates the process of unbalanced growth leading to cell death through the loss of normal assembly, function, or integrity of biomembranes. - Part of this work has been reported in preliminary form (CULBERTSON and HENRY 1974).


Th1 and Innate Lymphoid Cells Accumulate in Primary Sclerosing Cholangitis-associated Inflammatory Bowel Disease.
Primary sclerosing cholangitis [PSC] is an idiopathic chronic disorder of the hepatobiliary system associated with inflammatory bowel disease [IBD], mainly ulcerative colitis [UC]. Colitis in patients with PSC and UC [PSC-UC] exhibits characteristic features and is linked to increased colon cancer risk. Genetic studies have identified immune-related susceptibility genes that only partially overlap with those involved in IBD. These observations suggest that PSC-UC may represent a distinct form of IBD. It remains to be elucidated whether different immune mechanisms are involved in colitis in these patients. We aimed to evaluate systemic and intestinal T cell and innate lymphoid cell [ILC] responses, previously associated with IBD, in patients with PSC-UC compared with patients with UC and healthy controls. Blood samples and colorectal biopsies were collected from patients with PSC-UC, patients with UC, and healthy controls. T cell and ILC phenotypes were analysed by multicolour flow cytometry. Chemokine receptor [CCR] profiling of circulating T cells showed decreased CCR6-CXCR3+ Th1 cells in PSC-UC, but increased CCR6-CCR4+ Th2 cells only in UC, whereas increased CCR6+CCR4+ Th17 cells were found in both patient groups compared with healthy controls. Increased frequencies of IFN-γ secreting T cells were found in the colon of patients with PSC-UC compared with UC. Interestingly, we observed accumulation of ILC in the colon in PSC-UC. Our study suggests that PSC-UC represents a different immunological disorder from UC, characterised by increased intestinal Th1 and ILC responses. These results provide further evidence that PSC-UC may represent a distinct form of IBD.


Automated wide-field malaria parasite infection detection using Fourier ptychography on stain-free thin-smears.
Diagnosis of malaria in endemic areas is hampered by the lack of a rapid, stain-free and sensitive method to directly identify parasites in peripheral blood. Herein, we report the use of Fourier ptychography to generate wide-field high-resolution quantitative phase images of erythrocytes infected with malaria parasites, from a whole blood sample. We are able to image thousands of erythrocytes (red blood cells) in a single field of view and make a determination of infection status of the quantitative phase image of each segmented cell based on machine learning (random forest) and deep learning (VGG16) models. Our random forest model makes use of morphology and texture based features of the quantitative phase images. In order to label the quantitative images of the cells as either infected or uninfected before training the models, we make use of a Plasmodium berghei strain expressing GFP (green fluorescent protein) in all life cycle stages. By overlaying the fluorescence image with the quantitative phase image we could identify the infected subpopulation of erythrocytes for labelling purposes. Our machine learning model (random forest) achieved 91% specificity and 72% sensitivity while our deep learning model (VGG16) achieved 98% specificity and 57% sensitivity. These results highlight the potential for quantitative phase imaging coupled with artificial intelligence to develop an easy to use platform for the rapid and sensitive diagnosis of malaria.


Developmental shifts in children's sensitivity to visual speech: a new multimodal picture-word task.
This research developed a multimodal picture-word task for assessing the influence of visual speech on phonological processing by 100 children between 4 and 14 years of age. We assessed how manipulation of seemingly to-be-ignored auditory (A) and audiovisual (AV) phonological distractors affected picture naming without participants consciously trying to respond to the manipulation. Results varied in complex ways as a function of age and type and modality of distractors. Results for congruent AV distractors yielded an inverted U-shaped function with a significant influence of visual speech in 4-year-olds and 10- to 14-year-olds but not in 5- to 9-year-olds. In concert with dynamic systems theory, we proposed that the temporary loss of sensitivity to visual speech was reflecting reorganization of relevant knowledge and processing subsystems, particularly phonology. We speculated that reorganization may be associated with (a) formal literacy instruction and (b) developmental changes in multimodal processing and auditory perceptual, linguistic, and cognitive skills.


Geriatric grand rounds: hearing impairment and functional independence.
Hearing loss is a commonly encountered impairment in the elderly. It can profoundly affect physical, psychosocial and cognitive function. In addition to the actual hearing loss, other factors contribute to the hearing handicap experienced by older persons. These include difficulty in understanding speech in noisy situations, difficulty in supplementing hearing with visual information, and the slowing of cognitive and psychologic processes. Many forms of amplification are available and can be very useful in reversing much of the functional impairment associated with hearing loss. A comprehensive audiological assessment should be recommended before selecting the type of hearing aid or assistive listening device. Such a referral also affords an opportunity for more extensive aural rehabilitation.


Mechanical Compression Regulates Brain Cancer Cell Migration Through MEK1/Erk1 Pathway Activation and GDF15 Expression.
Mechanical compression is a common abnormality of brain tumors that has been shown to be responsible for the severe neurological defects of brain cancer patients representing a negative prognostic factor. Indeed, it is of note that patients that undergo resection exhibited higher survival rates than those subjected to biopsy only, suggesting that compressive forces generated during brain tumor growth play a key role in tumor progression. Despite the importance of mechanical compression in brain tumors, there is a lack of studies examining its direct effects on brain cancer cells and the mechanisms involved. In the present study, we used two brain cancer cell lines with distinct metastatic potential, the less aggressive H4 and the highly aggressive A172 cell lines, in order to study the effect of compression on their proliferative and migratory ability. Specifically, we used multicellular tumor spheroids (MCS) embedded in agarose matrix to show that compression strongly impaired their growth. Using mathematical modeling, we estimated the levels of compressive stress generated during the growth of brain MCS and then we applied the respective stress levels on brain cancer cell monolayers using our previously established transmembrane pressure device. By performing a scratch assay, we found that compression strongly induced the migration of the less aggressive H4 cells, while a less pronounced effect was observed for A172 cells. Analysis of the gene expression profile of both cell lines revealed that GDF15 and small GTPases are strongly regulated by mechanical compression, while GDF15 was further shown to be necessary for cells to migrate under compression. Through a phospho-proteomic screening, we further found that compressive stimulus is transmitted through the MEK1/Erk1 signaling pathway, which is also necessary for the migration of brain cancer cells. Finally, our results gave the first indication that GDF15 could regulate and being regulated by MEK1/Erk1 signaling pathway in order to facilitate the compression-induced brain cancer cell migration, rendering them along with small GTPases as potential targets for future anti-metastatic therapeutic innovations to treat brain tumors.


Adenovirus-mediated p53 gene transduction inhibits telomerase activity independent of its effects on cell cycle arrest and apoptosis in human pancreatic cancer cells.
Evidence for a relationship between overexpression of wild-type p53 and telomerase activity remains controversial. We investigated whether p53 gene transduction could cause telomerase inhibition in pancreatic cancer cell lines, focusing on the relation of transduction to growth arrest, cell cycle arrest, and apoptotic cell death. The cells were infected with recombinant adenovirus expressing wild-type p53 or p21WAF1 at a multiplicity of infection of 100 or were continuously exposed to 10 microM VP-16, which is well known to induce apoptosis. Adenovirus-mediated p53 gene transduction caused G1 cell cycle arrest, apoptosis, and resultant growth inhibition in MIA PaCa-2 cells; the cell number 2 days after infection was 50% of preinfection value, and 13% of the cells were dead. Moreover, the transduction resulted in complete depression of telomerase activity through down-regulation of hTERT mRNA expression. In contrast, p21WAF1 gene transduction only arrested cell growth and cell cycle at G1 phase, and VP-16 treatment inhibited cell growth with G2-M arrest and apoptosis; after treatment, the cell number was 73% of pretreatment, and 12% of the cells were dead. Neither p21WAF1 gene transduction nor VP-16 treatment caused telomerase inhibition. Similar results were obtained in two other pancreatic cancer cell lines, SUIT-2 and AsPC-1. Thus, our results demonstrate that the p53 gene transduction directly inhibits telomerase activity, independent of its effects on cell growth arrest, cell cycle arrest, and apoptosis.


The putative P-gp inhibitor telmisartan does not affect the transcellular permeability and cellular uptake of the calcium channel antagonist verapamil in the P-glycoprotein expressing cell line MDCK II MDR1.
Verapamil is used in high doses for the treatment of cluster headache. Verapamil has been described as a P-glycoprotein (P-gp, ABCB1) substrate. We wished to evaluate in vitro whether co administration of a P-gp inhibitor with verapamil could be a feasible strategy for increasing CNS uptake of verapamil. Fluxes of radiolabelled verapamil across MDCK II MDR1 monolayers were measured in the absence and presence of the putative P-gp inhibitor telmisartan (a clinically approved drug compound). Verapamil displayed a vectorial basolateral-to-apical transepithelial efflux across the MDCK II MDR1 monolayers with a permeability of 5.7 × 10(-5) cm sec(-1) compared to an apical to basolateral permeability of 1.3 × 10(-5) cm sec(-1). The efflux could be inhibited with the P-gp inhibitor zosuquidar. Zosuquidar (0.4 μmol/L) reduced the efflux ratio (PB-A/PA-B) for verapamil 4.6-1.6. The presence of telmisartan, however, only caused a slight reduction in P-gp-mediated verapamil transport to an efflux ratio of 3.4. Overall, the results of the present in vitro approach indicate, that clinical use of telmisartan as a P-gp inhibitor may not be an effective strategy for increasing brain uptake of verapamil by co-administration with telmisartan.


[The third generation vaccines].
The introduction of a new Hepatitis B-vaccine as the first licensed vaccine which is manufactored by gentechnological methods has encouraged generally to discuss advantages and problems of vaccines of the "Third Generation".


The influence of surface texture and wettability on initial bacterial adhesion on titanium and zirconium oxide dental implants.
This study aims to investigate bacterial adhesion on different titanium and ceramic implant surfaces, to correlate these findings with surface roughness and surface hydrophobicity, and to define the predominant factor for bacterial adhesion for each material. Zirconia and titanium specimens with different surface textures and wettability (5.0 mm in diameter, 1.0 mm in height) were prepared. Surface roughness was measured by perthometer (R a ) and atomic force microscopy, and hydrophobicity according to contact angles by computerized image analysis. Bacterial suspensions of Streptococcus sanguinis and Staphylococcus epidermidis were incubated for 2 h at 37 °C with ten test specimens for each material group and quantified with fluorescence dye CytoX-Violet and an automated multi-detection reader. Variations in surface roughness (R a ) did not lead to any differences in adhering S. epidermidis, but higher R a resulted in increased S. sanguinis adhesion. In contrast, higher bacterial adhesion was observed on hydrophobic surfaces than on hydrophilic surfaces for S. epidermidis but not for S. sanguinis. The potential to adhere S. sanguinis was significantly higher on ceramic surfaces than on titanium surfaces; no such preference could be found for S. epidermidis. Both surface roughness and wettability may influence the adhesion properties of bacteria on biomaterials; in this context, the predominant factor is dependent on the bacterial species. Wettability was the predominant factor for S. epidermidis and surface texture for S. sanguinis. Zirconia did not show any lower bacterial colonization potential than titanium. Arithmetical mean roughness values R a (measured by stylus profilometer) are inadequate for describing surface roughness with regard to its potential influence on microbial adhesion.


Effect of sodium loading and depletion on vascular reactivity and prostacyclin generation.
In order to estimate the modulatory activity of Prostacyclin (PGI2) to the vascular reactivity, pressor responses to angiotensin II (A II) and noradrenaline (NA) and PGI2 generation aorta were measured using the rats of sodium loading or of sodium depletion. On the sodium loading, the blood pressure increased gradually and plasma renin activity (PRA) and plasma aldosterone concentration (PAC) were decreased. The pressor responses to A II and NA were enhanced by sodium loading. The PGI2 generation of aorta was enhanced at the initial stage of sodium loading and decreased thereafter. The increased generation of PGI2 may represent the adaptive mechanism for the attenuation of the sustained elevation in blood pressure. It sodium depletion, the pressor responses to A II and NA were decreased, and PRA and PAC were elevated. PGI2 generation of aorta was also increased. These findings suggested that PGI2 could participate in blood pressure control mechanism on sodium loading and depletion.


Effect of vesicular stomatitis virus and Semliki Forest Virus on uptake of nutrients and intracellular cation concentration.
BHK-21 cells showed an increased ability to concentrate 2-deoxy-D-glucose (dGlc) 2 to 3 h after infection with vesicular stomatitis virus (VSV) or Semliki Forest virus (SFV), which began to be released at 2 and 3 h post-infection respectively; uptake of other nutrients was not affected in this way. Intracellular Na+ was either unchanged (VSV-infected cells) or increased (SFV-infected cells); K+ content was unchanged. These results do not support the current hypothesis that a non-specific increase in membrane permeability occurs in cells infected with rhabdoviruses or togaviruses.


10.4 kW coherently combined ultrafast fiber laser.
An ultrafast laser delivering 10.4 kW average output power based on a coherent combination of 12 step-index fiber amplifiers is presented. The system emits close-to-transform-limited 254 fs pulses at an 80 MHz repetition rate, and has a high beam quality (M2≤1.2) and a low relative intensity noise of 0.56% in the frequency range of 1 Hz to 1 MHz. Automated spatiotemporal alignment allows for hands-off operation.


Childhood mortality, family size and birth order in pre-industrial Europe.
Based on parish registers, demographic histories of Crulai (France), Tourouvre-au-Perche (France), and Geneva (Swizertland) established the childhood mortality experienced by complete sibships during periods of at least half a century before the Fench revolution. These observations may be presented as frequenceis in incomplete five-dimensional contingency tables. The five dimensions are: survival (living or dead), completed sibship size, birth order, type of family (according to completeness of information about family), and epoch (period in which the family lived). This paper reanalyzes these published data, using hierarchical log-linear models to discern which interations among the five variables can justifiably be inferred from the data. The neonatal and infant mortality rates of firstborn are probably higher than those of later sibs (in Crulai and Tourouvre). But mortality by age 20 (in Geneva) is associated strongly with the epoch, type of family, and family size, and not significantly with birth order. The increase in mortality with completed family size is insufficient to select, in an evolutionary sense, for limited family size.


The dilemma of science and morals.
The conflicts between science and morals which still continue to arise despite the apparent hegemony of atheistic scientism over traditional Judeo-Christianity in the twentieth century reflect a basic contradiction in the metaphysical foundation of Western lives. As was set forth by Machiavelli, the contradiction inherent in Western ethics is that it is based on the simultaneous belief in both objectively valid moral truths and purely relative values of communal purpose. The achievements of twentieth century science have intensified these contradictions. Modern physics has put in question the validity of its own metaphysical basis, namely the belief in Natural Law, and modern biology has been unable to come to terms with the Cartesian dualism of body and soul. By contrast, Chinese lives are comparatively free of these contradictions, being founded on the philosophies of Confucianism and Taoism, to which the concepts of objectively valid truth or Natural Law are foreign. Recent developments in Western attitudes regarding science and morals can be interpreted as a movement away from the traditional belief in absolute truths towards a Chinese relativism.


Rapid dissolution of DNA in a novel bio-based ionic liquid with long-term structural and chemical stability: successful recycling of the ionic liquid for reuse in the process.
DNA from salmon testes was solubilised in two bio-based ionic liquids up to 3.5% w/w in 6 h. No structural degradation of the molecule was observed for the sample solubilised in choline-indole-3-acetate (chol-IAA). However, the molecule was found to be denatured in choline-indole-3-butyrate (chol-IBA). The structural and chemical stability of the DNA molecules after six months of storage in the former was established. Further recyclability of the ionic liquid with very high yield (90-95%) for consecutive reuse in the redissolution of DNA was demonstrated.


Hepatic ultrastructure in congenital syphilis.
The ultrastructural changes in the liver in a case of congenital syphilis were studied. Treponema organisms were easily demonstrated in 1-micrometer sections stained by toluidine blue, as well as in electron micrographs. Most organisms were extracellular, but some were identified within hepatocytes.


Identification of methanol-soluble compounds in sesame and evaluation of antioxidant potential of its lignans.
The methanol extract of sesame (Sesamum indicum) seeds was fractionated and purified with the assistance of conventional column chromatography to afford 29 compounds including seven furofuran lignans. Among these isolates, (+)-samin (1) was obtained from the natural source for the first time. In addition, (-)-asarinin (30) and sesamol (31) were generated by oxidative derivation from (+)-sesamolin (2) and (+)-sesamin (3), two abundant lignans found in sesame seeds. To evaluate their in vitro antioxidant potential, the seven isolated lignans (1-7) and the two derivatives (30 and 31) were examined for the scavenging activities on DPPH free radicals and superoxide anions. Moreover, the capability of chelating ferrous ions and reducing power of these sesame lignans were also measured. The results suggest that, besides the well-known sesamolin and sesamin, the minor sesame lignans (+)-(7S,8'R,8R)-acuminatolide (5), (-)-piperitol (6), and (+)-pinoresinol (7) are also adequate active ingredients and may be potential sources for nutritional and pharmacological utilization.


Chemical Interaction of Hydrogen Radicals (H*) with Transition Metal Nitrides.
Transition metal nitrides (TMNs) are reported as protective coatings in reactive hydrogen environments. Although the permeation of H2 through TMN coatings is well reported, their reducibility in H* environments is less investigated. In this work, we categorize the interaction of H* with ambient exposed TiN, ZrN, HfN, VN, NbN, and TaN thin films at 700 °C into three classes. We find that in TiN and VN samples, H*-induced reduction was limited to the surface (≈ top 2 nm). Significant denitridation was observed in ZrN and HfN samples beneath the surface, along with an increase in the transition metal oxide (TMOx) fraction. Denitridation was observed in NbN and TaN samples as well, but the increase in the TMOx content was less than for ZrN and HfN. We propose a model in three steps: hydrogenation, formation of volatile species, and diffusion of subsurface atoms to the surface. We show that the interaction of H* with TiN, ZrN, HfN, VN, NbN, and TaN with partially oxidized surfaces can be explained using the preferred hydrogenation pathway (based on the work functions) and the thermodynamic driver for forming volatile species (NH3 and H2O; based on the change in Gibbs free energy).


Bisphenol A (BPA) and bisphenol S (BPS) alter the promoter activity of the ABCB1 gene encoding P-glycoprotein in the human placenta in a haplotype-dependent manner.
Exposure to bisphenols (BPA and BPS) during pregnancy can significantly affect fetal development and increase risk of adverse health consequences, however the underlying mechanisms are not fully elucidated. In human placenta, the efflux transporter P-glycoprotein (P-gp), encoded by the ABCB1 gene, extrudes its substrates from the trophoblasts back into the maternal circulation. Alterations in levels of placental P-gp could therefore significantly affect fetal exposure to xenobiotics that are P-gp substrates. The ABCB1 promoter contains many single nucleotide polymorphisms (SNPs). In the genome, SNPs are not arrayed as independent variants but as combinations forming defined haplotypes. Recently, we determined the haplotype sequences encompassing the ABCB1 promoter SNPs and found that promoter haplotypes differentially affect ABCB1 promoter activity. Here we investigate the effect of BPA and BPS on ABCB1 promoter activity by testing the hypothesis that BPA and BPS exposure affect ABCB1 promoter activity in a haplotype-dependent manner. Our data indicate that acute exposure to 50 nM BPA induced a significant haplotype-dependent increase in ABCB1 promoter activity (P < .05). However, acute exposure to 0.5 nM BPS induced a significant decrease (P < .05) in promoter activity that was haplotype-dependent. Chronic exposure to BPA and BPS individually (5 nM and 0.3 nM, respectively) or as a mixture (5 nM BPA:1.5 nM BPS) induced significant haplotype-dependent increases (P < .01) in ABCB1 promoter activity. Our data indicate that BPA and BPS significantly alter ABCB1 promoter activity in a haplotype- and exposure type- dependent manners. Such alteration could significantly impact placental P-gp levels and alter fetal exposure to many therapeutic and environmental xenobiotics.


[Resection of the saphenofemoral junction, when and why?].
Indications for 'simple resection of the saphenofemoral junction' are rare. Apart from second operations following incomplete saphenectomies, they are limited to the fairly exceptional cases of ostial incompetence involving the saphenous axis. But any correct saphenectomy will include the resection of the sephenofemoral junction. This is practised where there is an open saphenofemoral junction, and in clear cases of incompetence in the saphenous axis. Apart from these classic indications, resection of the (long) saphenofemoral junction is carried out in cases of 'aesthetic saphenectomy', as it avoids scars disfiguring the thigh or leg.


Long-term glucocorticoid levels measured in hair in patients with depressive and anxiety disorders.
Depressive and anxiety disorders have been linked to a dysregulated hypothalamus-pituitary-adrenal (HPA)-axis. Hair cortisol levels (HairF) reflect integrated long-term cortisol regulation and are therefore promising endocrine markers of chronic (psychological and physical) stress. Our aim was to assess hair cortisol levels in persons with a depressive and/or anxiety disorder and to compare their levels with that of persons in remission and healthy controls. Data from 1166 participants of the Netherlands Study of Depression and Anxiety (NESDA) were used, including 266 participants with a recent (1-month) diagnosis of a depressive and/or anxiety disorder, 655 participants with a diagnosis in remission, and 245 healthy controls. HairF was measured in the proximal three cm of scalp hair, using LC-MS/MS. Compared to the healthy controls no differences on HairF or HairE levels were found for depressive and anxiety disorders alone. However the presence of a comorbid depressive and anxiety disorder was significantly associated with increased HairF levels (β = 0.07; p = .031), as was the severity of depressive symptoms (β = 0.06; p = .029), but no differences were found on HairE nor the HairF:HairE ratio. Persons with current diagnosis of comorbid depression and anxiety show moderately higher levels of cortisol than patients with only depression or anxiety, or patients in remission and healthy controls, which may be indicative of a chronic state of hyperactivation of the HPA axis.


Identification of Dementia in Recent Medicare Claims Data, Compared With Rigorous Clinical Assessments.
Medicare fee-for-service (FFS) claims data are increasingly leveraged for dementia research. Few studies address the validity of recent claim data to identify dementia, or carefully evaluate characteristics of those assigned the wrong diagnosis in claims. We used claims data from 2014 to 2018, linked to participants administered rigorous, annual dementia evaluations in 5 cohorts at the Rush Alzheimer's Disease Center. We compared prevalent dementia diagnosed through the 2016 cohort evaluation versus claims identification of dementia, applying the Bynum-standard algorithm. Of 1 054 participants with Medicare Parts A and B FFS in a 3-year window surrounding their 2016 index date, 136 had prevalent dementia diagnosed during cohort evaluations; the claims algorithm yielded 217. Sensitivity of claims diagnosis was 79%, specificity 88%, positive predictive value 50%, negative predictive value 97%, and overall accuracy 87%. White participants were disproportionately represented among detected dementia cases (true positive) versus cases missed (false negative) by claims (90% vs 75%, respectively, p = .04). Dementia appeared more severe in detected than missed cases in claims (mean Mini-Mental State Exam = 15.4 vs 22.0, respectively, p < .001; 28% with no limitations in activities of daily living versus 45%, p = .046). By contrast, those with "over-diagnosis" of dementia in claims (false positive) had several worse health indicators than true negatives (eg, self-reported memory concerns = 51% vs 29%, respectively, p < .001; mild cognitive impairment in cohort evaluation = 72% vs 44%, p < .001; mean comorbidities = 7 vs 4, p < .001). Recent Medicare claims perform reasonably well in identifying dementia; however, there are consistent differences in cases of dementia identified through claims than in rigorous cohort evaluations.


Monitored anesthesia care is not synonymous with low risk.
Monitored anesthesia care can lead operating room (OR) personnel, both perioperative nurses and anesthesia providers, into a sense of complacency. It is not general anesthesia, so there is no stress surrounding intubation and extubation and the patient is in control of his/her own respirations. When used properly, narcotics and sedative drugs sedate and relieve pain, but should not lead to respiratory compromise. As a result, circumstances should be under control when competent staff are present. However, because unanticipated events do happen even under the most benign-appearing occurrences, there must be a sense of vigilance by everyone in the OR as this case study relates.


Microbial biofilms in seafood: a food-hygiene challenge.
Seafood forms a part of a healthy diet. However, seafood can be contaminated with foodborne pathogens, resulting in disease outbreaks. Because people consume large amounts of seafood, such disease outbreaks are increasing worldwide. Seafood contamination is largely due to the naturally occurring phenomenon of biofilm formation. The common seafood bacterial pathogens that form biofilms are Vibrio spp., Aeromonas hydrophila, Salmonella spp., and Listeria monocytogenes. As these organisms pose a global health threat, recent research has focused on elucidating methods to eliminate these biofilm-forming bacteria from seafood, thereby improving food hygiene. Therefore, we highlight recent advances in our understanding of the underlying molecular mechanisms of biofilm formation, the factors that regulate biofilm development and the role of quorum sensing and biofilm formation in the virulence of foodborne pathogens. Currently, several novel methods have been successfully developed for controlling biofilms present in seafood. In this review, we also discuss the epidemiology of seafood-related diseases and the novel methods that could be used for future control of biofilm formation in seafood.


Measuring Healthcare Providers' Performances Within Managed Competition Using Multidimensional Quality and Cost Indicators.
The Dutch healthcare system is in transition towards managed competition. In theory, a system of managed competition involves incentives for quality and efficiency of provided care. This is mainly because health insurers contract on behalf of their clients with healthcare providers on, potentially, quality and costs. The paper develops a strategy to comprehensively analyse available multidimensional data on quality and costs to assess and report on the relative performance of healthcare providers within managed competition. We had access to individual information on 2409 clients of 19 Dutch diabetes care groups on a broad range of (outcome and process related) quality and cost indicators. We carried out a cost-consequences analysis and corrected for differences in case mix to reduce incentives for risk selection by healthcare providers. There is substantial heterogeneity between diabetes care groups' performances as measured using multidimensional indicators on quality and costs. Better quality diabetes care can be achieved with lower or higher costs. Routine monitoring using multidimensional data on quality and costs merged at the individual level would allow a systematic and comprehensive analysis of healthcare providers' performances within managed competition.


Dark chocolate as a stable carrier of microencapsulated Akkermansia muciniphila and Lactobacillus casei.
The viability of probiotics is affected by several factors during manufacturing, storage and gastrointestinal tract passage. Protecting the probiotics from harmful conditions is particularly critical for oxygen sensitive species like Akkermansia muciniphila, a bacterium which recently has been proposed as a next-generation probiotic candidate. Previously, we have developed a protocol for microencapsulating A. muciniphila in a xanthan/gellan gum matrix. Here, we report the enhanced survival during storage and in vitro gastric passage of microencapsulated A. muciniphila embedded in dark chocolate. Lactobacillus casei, as a representative species of traditional probiotics, was included in order to compare its behavior with that of A. muciniphila. For A. muciniphila we observed a 0.63 and 0.87 log CFU g-1 reduction during 60 days storage at 4°C or 15°C, respectively. The viability of L. casei remained stable during the same period. During simulated gastric transit (pH 3), microencapsulated A. muciniphila embedded in chocolate showed 1.80 log CFU mL-1 better survival than naked cells, while for L. casei survival was improved with 0.8 log CFU mL-1. In a hedonic sensory test, dark chocolate containing microcapsules were not significantly different from two commercially available chocolates. The developed protocol constitutes a promising approach for A. muciniphila dosage.


Chloroplast biogenesis 87: Evidence of resonance excitation energy transfer between tetrapyrrole intermediates of the chlorophyll biosynthetic pathway and chlorophyll a.
The thorough understanding of photosynthetic membrane assembly requires a deeper knowledge of the coordination of chlorophyll (Chl) and thylakoid apoprotein biosynthesis. As a working model for future investigations, we have proposed three Chl-thylakoid apoprotein biosynthesis models, namely, a single-branched Chl biosynthetic pathway (SBP) single-location model, an SBP multilocation model and a multibranched Chl biosynthetic pathway (MBP) sublocation model. Rejection or validation of these models can be probed by determination of resonance excitation energy transfer between various tetrapyrrole intermediates of the Chl biosynthetic pathway and various thylakoid Chl-protein complexes. In this study we describe the detection of resonance energy transfer between protoporphyrin IX (Proto), Mg-Proto and its monomethyl ester (Mp(e)) and divinyl and monovinyl protochlorophyllide a (Pchlide a) and several Chl-protein complexes. Induction of various amounts of tetrapyrrole accumulation in green photoperiodically grown cucumber cotyledons and barley leaves was achieved by dark incubation of excised tissues with delta-aminolevulinic acid (ALA) and various concentrations of 2,2'-dipyridyl for various periods of time. Controls were incubated in distilled water. After plastid isolation, treated and control plastids were diluted in buffered glycerol to the same Chl concentration. Excitation spectra were then recorded at 77 K at emission maxima of about 686, 694 and 738 nm. Resonance excitation energy transfer from Proto, Mp(e) and Pchlide a to Chl-protein complexes emitting at 686, 694 and 738 nm was observed by calculation of treated minus control difference excitation spectra. The occurrence of resonance excitation energy transfer between anabolic tetrapyrroles and Chl-protein complexes appeared as well-defined excitation bands with excitation maxima corresponding to those of Proto, Mp(e) and Pchlide a. Furthermore, it appeared that resonance excitation energy transfer from multiple short-wavelength, medium-wavelength and long-wavelength Proto, Mp(e) and Chlide a sites to various Chl-protein complexes took place. Because resonance excitation transfer from donors to acceptors cannot take place at distances larger than 100 A, it is proposed that the observed resonance excitation energy transfers are not compatible with the SBP single-location Chl biosynthesis thylakoid membrane biogenesis model. The latter assumes that a single-branched Chl biosynthetic pathway located in the center of a 450 x 130 A photosynthetic unit generates all of the Chl needed for the assembly of all Chl-protein complexes.


Quality of life among patients with multiple sclerosis and voiding dysfunction: a cross-sectional study.
Evaluating the usefulness of treatment requires a direct measurement of the health-related quality of life (QOL). Therefore, this study was carried out aiming to determine the QOL of patients with MS and voiding dysfunction. This cross-sectional study was carried out using multi-stage random cluster sampling method on 602 patients with MS in Isfahan, Iran. All data were collected through interviews using standard questionnaires including International Prostate Symptom Score (IPSS), and the Multiple Sclerosis Quality of Life-54 (MSQOL-54). Data were analyzed using descriptive and inferential statistical tests. The prevalence rate of mixed, irritative, and obstructive urinary symptoms was 52.2, 25.5, and 6.5%, respectively. The mixed symptom had the highest prevalence among men and women with rates of 56.5 and 51.1%, respectively. The prevalence of irritative and obstructive symptoms was, respectively, higher and statistically significant among women alone and men alone (P < 0.05). The prevalence of irritative symptoms was higher among patients with MS, EDSS score ≤ 3, disease duration of less than 5 years, and with clinically isolated syndrome. In addition, the prevalence of mixed symptoms was higher among patients with MS of over 30 years of age with a Pre-high school degree, severe disability, disease duration of over 10 years, and progressive MS; the difference was statistically significant (P < 0.05). There was a difference in the combined dimensions of physical and mental health of QOL between the two groups with and without urinary symptoms (P < 0.05). Logistic regression analysis revealed that there was a higher probability of a urinary problems among patients with MS and high age [3.273 (1.083-9.860); P = 0.035]. Mixed urinary symptoms are highly prevalent among MS patients and affect QOL dimensions. In order to improve QOL, more attention and focus should be paid to urinary problems in MS patients.


Small-bowel mucosal healing assessment by capsule endoscopy as a predictor of long-term clinical remission in patients with Crohn's disease: a systematic review and meta-analysis.
Capsule endoscopy (CE) may be used for the evaluation and follow-up of patients with Crohn's disease. CE scores correspond to the degree of mucosal inflammation, a major therapeutic target. The aim of this meta-analysis and systematic review was to determine whether mucosal healing assessment by CE may serve as a predictor of clinical remission in patients with Crohn's disease. To identify observational or controlled English-language full-text studies assessing mucosal healing by CE in patients with Crohn's disease up to 30 September 2016, we searched PubMed, Embase, Central, Medline, and Scopus using the key words 'mucosal healing' and 'capsule endoscopy'. A meta-analysis was carried out using 'Comprehensive meta-analysis' software. Pooled odds ratios and 95% confidence intervals were calculated. Five observational studies including 142 patients from five countries fulfilled the inclusion criteria. No publication bias was found by funnel plot. The mucosal healing CE score was found to be significantly associated with improved outcome after a follow-up of 12 weeks to 24 months, with an odds ratio of 11.06 (95% confidence interval: 3.74-32.73, P<0.001). The degree of heterogeneity among the studies was small (Q=2.014, d.f.[Q]=3, P=0.569 and I=0). Endoscopy scores may play a role in the long-term prognostic evaluation of patients with Crohn's disease. Our results may be accepted as proof of concept, but larger studies are needed to corroborate these findings.


A single 36-h water-only fast vastly remodels the plasma lipidome.
Prolonged fasting, characterized by restricting caloric intake for 24 h or more, has garnered attention as a nutritional approach to improve lifespan and support healthy aging. Previous research from our group showed that a single bout of 36-h water-only fasting in humans resulted in a distinct metabolomic signature in plasma and increased levels of bioactive metabolites, which improved macrophage function and lifespan in C. elegans. This secondary outcome analysis aimed to investigate changes in the plasma lipidome associated with prolonged fasting and explore any potential links with markers of cardiometabolic health and aging. We conducted a controlled pilot study with 20 male and female participants (mean age, 27.5 ± 4.4 years; mean BMI, 24.3 ± 3.1 kg/m2) in four metabolic states: (1) overnight fasted (baseline), (2) 2-h postprandial fed state (fed), (3) 36-h fasted state (fasted), and (4) 2-h postprandial refed state 12 h after the 36-h fast (refed). Plasma lipidomic profiles were analyzed using liquid chromatography and electrospray ionization mass spectrometry. Several lipid classes, including lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), phosphatidylethanolamine, and triacylglycerol were significantly reduced in the 36-h fasted state, while free fatty acids, ceramides, and sphingomyelin were significantly increased compared to overnight fast and fed states (P < 0.05). After correction for multiple testing, 245 out of 832 lipid species were significantly altered in the fasted state compared to baseline (P < 0.05). Random forest models revealed that several lipid species, such as LPE(18:1), LPC(18:2), and FFA(20:1) were important features in discriminating the fasted state from both the overnight fasted and postprandial state. Our findings indicate that prolonged fasting vastly remodels the plasma lipidome and markedly alters the concentrations of several lipid species, which may be sensitive biomarkers of prolonged fasting. These changes in lipid metabolism during prolonged fasting have important implications for the management of cardiometabolic health and healthy aging, and warrant further exploration and validation in larger cohorts and different population groups.


Formation of a ternary complex: actin, 5'-adenylyl imidodiphosphate, and the subfragments of myosin.
The formation of the ternary complex composed of actin, 5'-adenylyl imidodiphosphate [AMP-P(NH)P], and myosin subfragment 1 (S-1) was studied using the analytical ultracentrifuge with UV optics, which enabled the direct determination of the extent of dissociation of actin.S-1 (acto.S-1) by AMP-P(NH)P. In contrast to the reaction with ATP, at saturating levels of AMP-P(NH)P (1.5 mM), extensive formation of the ternary acto.S-1.AMP-P(NH)P complex occurs at 22 degrees . With 40 muM actin present, AMP-P(NH)P causes almost no dissociation of the acto.S-1 complex at 0.04 M ionic strength, while even at 0.22 M ionic strength one-third of the S-1 remains associated with actin and AMP-P(NH)P in a ternary complex. A detailed study of the binding of S-1.AMP-P(NH)P to actin using the Scatchard plot analysis shows that, at saturation, 1 mol of S-1.AMP-P(NH)P binds per mol of actin monomer. There appears to be no cooperativity occurring as the S-1.AMP-P(NH)P binds along the actin filament, with the possible exception of a slight positive cooperativity when most of the sites on the actin filament are saturated. The turbidity of the ternary complex is identical to the turbidity of acto.S-1 alone. Preliminary experiments with the two-headed subfragment of myosin, heavy meromyosin (HMM), show that the binding of HMM.[AMP-P(NH)P](2) to actin is only about twice as strong as the binding of S-1.AMP-P(NH)P to actin, indicating that the second head contributes very little to the free energy of binding.


Post-traumatic epilepsy: current and emerging treatment options.
Traumatic brain injury (TBI) leads to many undesired problems and complications, including immediate and long-term seizures/epilepsy, changes in mood, behavioral, and personality problems, cognitive and motor deficits, movement disorders, and sleep problems. Clinicians involved in the treatment of patients with acute TBI need to be aware of a number of issues, including the incidence and prevalence of early seizures and post-traumatic epilepsy (PTE), comorbidities associated with seizures and anticonvulsant therapies, and factors that can contribute to their emergence. While strong scientific evidence for early seizure prevention in TBI is available for phenytoin (PHT), other antiepileptic medications, eg, levetiracetam (LEV), are also being utilized in clinical settings. The use of PHT has its drawbacks, including cognitive side effects and effects on function recovery. Rates of recovery after TBI are expected to plateau after a certain period of time. Nevertheless, some patients continue to improve while others deteriorate without any clear contributing factors. Thus, one must ask, 'Are there any actions that can be taken to decrease the chance of post-traumatic seizures and epilepsy while minimizing potential short- and long-term effects of anticonvulsants?' While the answer is 'probably,' more evidence is needed to replace PHT with LEV on a permanent basis. Some have proposed studies to address this issue, while others look toward different options, including other anticonvulsants (eg, perampanel or other AMPA antagonists), or less established treatments (eg, ketamine). In this review, we focus on a comparison of the use of PHT versus LEV in the acute TBI setting and summarize the clinical aspects of seizure prevention in humans with appropriate, but general, references to the animal literature.


Raddeanin A suppresses lung cancer cell proliferation via induction of apoptosis and increased production of ROS.
At present, in vitro cell experiments have confirmed that RaddeaninA can effectively inhibit the proliferation of some tumor cells, but the effect of RaddeaninA on lung cancer cells has not been observed. Therefore, this study explored its effect on lung cancer cells and its mechanism of action. Human lung cancer cell lines were treated with serum-free medium and varied concentrations of Raddeanin A. Cell proliferation and apoptosis were determined using MTT, and flow cytometric assays, respectively. The intracellular level of ROS was determined using DCFH-DA assay. Protein and mRNA expressions of bax, bcl-2 and cyt c were measured using Western blotting and qRT-PCR. RaddeaninA treatment can promote PC-9 cell apoptosis in a time and dose-dependent manner (p<0.05). Treatment of PC-9 cells with Raddeanin significantly and dose-dependently increased the activities of caspase-9 and caspase-3 (p<0.05), and led to significant and dose-dependent increases in ROS levels (p<0.05). Treatment of PC-9 cells with Raddeanin A led to significant and dose-dependent decreases in mitochondrial membrane potential (p<0.05). It significantly and dose-dependently upregulated bax mRNA and protein expressions, but down-regulated bcl-2 mRNA and protein expressions significantly and dose-dependently (p<0.05). On the other hand, Raddeanin significantly and dose-dependently down-regulated cytoplasmic bax protein expression, while upregulating cyt c expression (p<0.05). Similarly, bax protein expression was significantly and dose-dependently upregulated in mitochondria, but the corresponding cyt c expression was significantly and dose-dependently down-regulated (p<0.05). Raddeanin A is a potential and effective lung cancer chemotherapy drug, which can induce lung cancer cell apoptosis and inhibit proliferation.


Circadian rhythms and metabolic syndrome: from experimental genetics to human disease.
The incidence of the metabolic syndrome represents a spectrum of disorders that continue to increase across the industrialized world. Both genetic and environmental factors contribute to metabolic syndrome and recent evidence has emerged to suggest that alterations in circadian systems and sleep participate in the pathogenesis of the disease. In this review, we highlight studies at the intersection of clinical medicine and experimental genetics that pinpoint how perturbations of the internal clock system, and sleep, constitute risk factors for disorders including obesity, diabetes mellitus, cardiovascular disease, thrombosis and even inflammation. An exciting aspect of the field has been the integration of behavioral and physiological approaches, and the emerging insight into both neural and peripheral tissues in disease pathogenesis. Consideration of the cell and molecular links between disorders of circadian rhythms and sleep with metabolic syndrome has begun to open new opportunities for mechanism-based therapeutics.


Global expression analysis of gene regulatory pathways during endocrine pancreatic development.
To define genetic pathways that regulate development of the endocrine pancreas, we generated transcriptional profiles of enriched cells isolated from four biologically significant stages of endocrine pancreas development: endoderm before pancreas specification, early pancreatic progenitor cells, endocrine progenitor cells and adult islets of Langerhans. These analyses implicate new signaling pathways in endocrine pancreas development, and identified sets of known and novel genes that are temporally regulated, as well as genes that spatially define developing endocrine cells from their neighbors. The differential expression of several genes from each time point was verified by RT-PCR and in situ hybridization. Moreover, we present preliminary functional evidence suggesting that one transcription factor encoding gene (Myt1), which was identified in our screen, is expressed in endocrine progenitors and may regulate alpha, beta and delta cell development. In addition to identifying new genes that regulate endocrine cell fate, this global gene expression analysis has uncovered informative biological trends that occur during endocrine differentiation.


The Synaptic Vesicle Glycoprotein 2: Structure, Function, and Disease Relevance.
The synaptic vesicle glycoprotein 2 (SV2) family is comprised of three paralogues: SV2A, SV2B, and SV2C. In vertebrates, SV2s are 12-transmembrane proteins present on every secretory vesicle, including synaptic vesicles, and are critical to neurotransmission. Structural and functional studies suggest that SV2 proteins may play several roles to promote proper vesicular function. Among these roles are their potential to stabilize the transmitter content of vesicles, to maintain and orient the releasable pool of vesicles, and to regulate vesicular calcium sensitivity to ensure efficient, coordinated release of the transmitter. The SV2 family is highly relevant to human health in a number of ways. First, SV2A plays a role in neuronal excitability and as such is the specific target for the antiepileptic drug levetiracetam. SV2 proteins also act as the target by which potent neurotoxins, particularly botulinum, gain access to neurons and exert their toxicity. Both SV2B and SV2C are increasingly implicated in diseases such as Alzheimer's disease and Parkinson's disease. Interestingly, despite decades of intensive research, their exact function remains elusive. Thus, SV2 proteins are intriguing in their potentially diverse roles within the presynaptic terminal, and several recent developments have enhanced our understanding and appreciation of the protein family. Here, we review the structure and function of SV2 proteins as well as their relevance to disease and therapeutic development.


Examining the effect of the dipole moment on charge separation in donor-acceptor polymers for organic photovoltaic applications.
A new low band gap copolymer PBB3 containing [6,6']bi[thieno[3,4-b]thiophenyl]-2,2'-dicarboxylic acid bis-(2-butyloctyl) ester (BTT) and 4,8-bis(2-butyloctyl)benzo[1,2-b:4,5-b']dithiophene (BDT) units was synthesized and tested for solar cell efficiency. PBB3 showed a broad absorbance in the near-IR region with a substantially red-shifted (by more than 100 nm) λ(max) at 790 nm as compared to the PTB series of polymers, which have been previously reported. The PBB3 polymer also showed both a favorable energy level match with PCBM (with a LUMO energy level of -3.29 eV) and a favorable film domain morphology as evidenced by TEM images. Despite these seemingly optimal parameters, a bulk heterojunction (BHJ) photovoltaic device fabricated from a blend of PBB3 and PC(71)BM showed an overall power conversion efficiency (PCE) of only 2.04% under AM 1.5G/100 mW cm(-2). The transient absorption spectra of PBB3 showed the absence of cationic and pseudo charge transfer states that were observed previously in the PTB series polymers, which were also composed of alternating thienothiophene (TT) and BDT units. We compared the spectral features and electronic density distribution of PBB3 with those of PTB2, PTB7, and PTBF2. While PTB2 and PTB7 have substantial charge transfer characteristics and also relatively large local internal dipoles through BDT to TT moieties, PTBF2 and PBB3 have minimized internal dipole moments due to the presence of two adjacent TT units (or two opposing fluorine atoms in PTBF2) with opposite orientations or internal dipoles. PBB3 showed a long-lived excitonic state and the slowest electron transfer dynamics of the series of polymers, as well as the fastest recombination rate of the charge-separated (CS) species, indicating that electrons and holes are more tightly bound in these species. Consequently, substantially lower degrees of charge separation were observed in both PBB3 and PTBF2. These results show that not only the energetics but also the internal dipole moment along the polymer chain may be critical in maintaining the pseudocharge transfer characteristics of these systems, which were shown to be partially responsible for the high PCE device made from the PTB series of low band gap copolymers.


Evaluation of bone mineral density in adolescent idiopathic scoliosis using a three-dimensional finite element model: a retrospective study.
Adolescent idiopathic scoliosis (AIS) is often accompanied by osteopenia and osteoporosis, which can cause serious complications. The aim of this study was to determine the specific bone mineral density (BMD) of each vertebral body in patients with AIS using biomechanical finite element modeling based on three-dimensional (3D) reconstruction. This retrospective study involved 56 patients with AIS. Computed tomography (CT) and radiography were performed. Spinal vertebrae were segmented from the spinal CT images of patients with AIS to reconstruct 3D vertebral models. The vertebral models were meshed into tetrahedral finite elements to assess the BMD. The mean main curve Cobb angle was 88.6 ± 36.7°, and the mean kyphosis angle was 36.8 ± 31.5°. The mean BMD of the global spine was 0.83 ± 0.15 g/cm2. The highest BMD was measured on the concave side of the apex (0.98 ± 0.16 g/cm2). Apical vertebral BMD was negatively correlated with age and height (r = - 0.490, p = 0.009 and r = - 0.478, p = 0.043, respectively). There were no significant differences in BMD values between the concave and convex sides (p > 0.05). The 3D finite element modeling of BMD in patients with AIS is a reliable and accurate BMD measurement method. Using this method, the overall BMD of patients with AIS was shown to gradually decrease from the top to the bottom of the spine. Our findings provide valuable insights for surgical planning, choice of screw trajectories, and additional biomechanical analyzes using finite element models in the context of scoliosis.


Adult acute lymphoblastic leukemia phenotypes defined by monoclonal antibodies.
Pretreatment peripheral blood and/or bone marrow blasts from 90 adults with acute lymphoblastic leukemia (ALL) were analyzed as part of a prospective treatment protocol study. Specimens were tested by immunofluorescence cytofluorometry for reactivity with the following monoclonal antibodies (MoAbs): BA-1 (B cell antigen); T101, OKT11 (pan-T cell antigens [T]); 3A1 (T cell antigen); MCS-2 (myeloid antigen); J5 common ALL antigen (CALLA); BA4 (Ia antigen [Ia]); BA-2 (lymphohematopoietic antigen). Four major phenotypic groups were identified: B lineage ALL (BA-1+T-) (64%), T lineage ALL (T+BA-1-MCS-2-) (13%), unclassified ALL (BA-1-MCS-2-CALLA-T-) (9%) and myeloid antigen ALL (MCS-2+CALLA-T-) (7%). An additional group of patients, miscellaneous ALL (7%), was comprised of cases with unusual marker profiles. In B lineage ALL, all cases tested were Ia+MCS-2-, and the vast majority were CALLA+ (84%). In T lineage ALL, 42% expressed CALLA or Ia positivity. In unclassified ALL, the predominant phenotype was Ia+BA-2+. In myeloid antigen ALL, two of four tested were 3A1+ and all cases evaluated were BA-1-. Patients with myeloid antigen ALL were older (median age, 66 years) than patients in the other groups. The T lineage ALL group had higher leukocyte counts (median WBCs, 183,000/microL) and an increased incidence of anterior mediastinal mass at presentation. All patients received identical induction therapy. In CALLA+B lineage ALL, 30 of 46 (65%) achieved a complete remission. While the number of patients evaluated was small, 9 of 9 CALLA-B-lineage ALL and only two of six myeloid antigen ALL cases responded with a complete remission. The data suggest that these MoAbs are useful in the characterization of adult ALL.


Evidence that cold preservation-induced microcirculatory injury in liver allografts is not mediated by oxygen-free radicals or cell swelling in the rat.
The critical injury of cold preservation is to the hepatic microcirculation. Oxygen free radical injury and cell swelling have been proposed to be causes of allograft failure, and new solutions such as Marshall's isotonic citrate and University of Wisconsin (UW) solutions were designed to prevent cell swelling and free radical injury. Experiments were done to determine whether Marshall's solution and UW solution protect the microcirculation, and whether they do so by preventing cell swelling or free radical-induced injury. To determine if the new solutions reduce sinusoidal lining cell injury, rat livers were examined after preservation at 4 degrees C in NaCl 0.9% and CaCl2 2 mM for 4 hr and 8 hr, in Collins' solution for 8 hr, and in both UW and isotonic citrate solutions for 8 hr and 16 hr. Next, the role of cell swelling in preservation injury was studied by storing livers in hypotonic solutions that accelerate liver weight gain, and in a choline chloride-based preservation solution. Finally, to evaluate the role of active oxygen species, SOD, catalase, and allopurinol were added to preservation solutions. The effect of allopurinol alone was also studied. In a related study, sucrose was substituted for the free radical scavenger, mannitol, in isotonic citrate solution. All livers were studied by light microscopy after perfusion-fixation. Storage in UW and isotonic citrate solutions resulted in clear improvement in the morphology of the sinusoidal lining. Increasing the rate of liver weight gain by the use of hypotonic solutions did not accelerate the endothelial injury. Choline chloride-based solution prevented weight gain during preservation, but unlike UW or isocitrate solutions it did not retard the microcirculatory injury. After preservation in the presence of SOD and catalase, or allopurinol, no improvement in the defined morphological features of the endothelial injury was noted when compared with respective controls; nor was the benefit of isotonic citrate solution lessened by the removal of the free radical scavenger mannitol. We conclude that microvascular injury produced by cold injury is due neither to free radical-mediated injury nor to cell swelling. As both UW and isotonic citrate solutions provide significant protection to the microcirculation, they must do so by a yet-undetermined mechanism.


Relationship between anterior inferior cerebellar artery and facial-vestibulocochlear nerve complex: an anatomical and magnetic resonance images correlation study.
For the successful microneurosurgical treatment of CP angle located pathologies, an understanding of the relationship and variations between neural and vascular structures and a certain diagnosis are the most valuable factors for surgeons. CP angle lesions have now become a visible area by advances in magnetic resonance imaging (MRI) technology. An evaluation of this area and the decision for a neurosurgical decompression procedure are easier than before. Twenty unfixed adult human cadaver specimens, that have no sign of central nervous system pathology, were obtained and dissected bilaterally at routine autopsy. The facial-vestibulocochlear (VII - VIIIth) nerve complex and the anterior inferior cerebellar artery (AICA) were identified in all specimens. Thirteen of the 40 (32.5 %) AICA were situated ventrally and fourteen (35 %) were located dorsally to the VII - VIIIth nerve complex. Thirteen (32.5 %) passed between the VIIth and the VIIIth nerve fibers. Five of the 40 (12.5 %) AICA had a loop near the nerve complex and then passed the nerves ventrally or dorsally. In an MRI study 74 adult persons (148 sides) were investigated by using three-dimensional Fourier transformation constructive interference in the steady state technique (3D FT-CISS) on a 1.5 Tesla MRI system (Siemens Magnetom, Erlangen, Germany). The results were as follows; 48 AICA (32.4 % of all 148 AICA) were situated ventrally to the VII-VIIIth nerve complex, 45 AICA (30.4 %) were situated dorsally to the VII-VIIIth nerve complex, and the AICA passed between the VIIth and VIIIth nerves in 51 samples (34.5 %). In four of the 148 CP angles (2.7 %), the AICA was not identified. There was an AICA loop coursing to the internal acoustic meatus in 15 patients (10.1 %). In this study, we examined the relations between VIIth and VIIIth nerve complex and the AICA in cadaver and MRI materials for an understanding of the value and reliability of the radiological data. This study also shows the anatomical variation between these structures.


Application of rearrangements of T cell receptor gene in the diagnosis of mediastinal T-lymphoblastic lymphoma: a case report.
The cytologic diagnosis of lymphoid cells in pleural effusion is not readily apparent. The atypical lymphocytes in reactive process or viral infection may mimic immature lymphoblasts in lymphoblastic lymphoma. Therefore, an objective and reliable method to identify malignant lymphoid cells is important. Here we reported the application of T cell receptor (TCR) gene analysis, together with cytomorphology and immunophenotypic studies, to establish the diagnosis of T-lymphoblastic lymphoma in a 20-year-old female who presented with a mediastinal tumor and pleural effusion. The diagnosis of malignant lymphoma was suspected by cytology and then confirmed by TCR beta chain (TCR beta) gene rearrangement. The rearrangement of TCR beta gene revealed by Southern blot technique implied a T-monoclonal lymphoid population which is most likely malignant. Immunophenotypic studies revealed expression of TdT and T cell antigens indicating a T lymphoblastic lymphoma. As shown in this case, antigen receptor gene analysis can be conducted with a much smaller number of cells to identify a malignant clone among a heterogeneous cell population.


Atomic-scale control of graphene magnetism by using hydrogen atoms.
Isolated hydrogen atoms absorbed on graphene are predicted to induce magnetic moments. Here we demonstrate that the adsorption of a single hydrogen atom on graphene induces a magnetic moment characterized by a ~20-millielectron volt spin-split state at the Fermi energy. Our scanning tunneling microscopy (STM) experiments, complemented by first-principles calculations, show that such a spin-polarized state is essentially localized on the carbon sublattice opposite to the one where the hydrogen atom is chemisorbed. This atomically modulated spin texture, which extends several nanometers away from the hydrogen atom, drives the direct coupling between the magnetic moments at unusually long distances. By using the STM tip to manipulate hydrogen atoms with atomic precision, it is possible to tailor the magnetism of selected graphene regions.


Accelerating the Calculation of Protein-Ligand Binding Free Energy and Residence Times Using Dynamically Optimized Collective Variables.
Elucidation of the ligand/protein binding interaction is of paramount relevance in pharmacology to increase the success rate of drug design. To this end, a number of computational methods have been proposed; however all of them suffer from limitations since the ligand binding/unbinding transitions to the molecular target involve many slow degrees of freedom that hamper a full characterization of the binding process. Being able to express this transition in simple and general slow degrees of freedom would give a distinctive advantage, since it would require minimal knowledge of the system under study, while in turn it would elucidate its physics and accelerate the convergence speed of enhanced sampling methods relying on collective variables. In this study we pursuit this goal by combining for the first time variational approach to conformational dynamics with funnel metadynamics. In so doing, we predict for the benzamidine/trypsin system the ligand binding mode, and we accurately compute the absolute protein-ligand binding free energy and unbinding rate at unprecedented low computational cost. Finally, our simulation protocol reveals the energetics and structural details of the ligand binding mechanism and shows that water and binding pocket solvation/desolvation are the dominant slow degrees of freedom.


Integrating psychobiography into psychology's mainstream: Introduction to the special section.
Psychobiography, the intense study of a particular person, has long existed at the fringes of mainstream psychology. This special section is comprised of two articles that take important steps to integrate psychobiography into the mainstream of psychology. Schultz and Lawrence (2017) do so by reviewing recent examples of how contemporary psychobiographers have used empirically supported methods and theories to reach conclusions about the individuals under study. Ponterotto and Reynolds (2017) do so by articulating important ethical issues relevant to the practice of psychobiography, a topic on which the American Psychological Association's current ethical code provides little guidance (American Psychological Association, 2017). This introduction closes by proposing that mainstream psychology might better appreciate how psychobiography contributes to psychological knowledge and practice if psychobiographers also clearly articulated their criteria for "good science," better explained how psychobiographical methods complement more mainstream methodological approaches, and empirically tested whether training in psychobiography helps clinicians better understand and treat their clients. (PsycINFO Database Record


Complex wound closure, excisional biopsy, and use of simple flaps.
The primary care physician can utilize the various techniques discussed in this article to close complex lacerations, modify scars, and perform excisional biopsies. Various methods for closure of square, triangular, semicircular, circular, and rectangular defects were described. The principle of flap advancement and Z-plasties was explained, and the method for performing excisional biopsies was detailed. These techniques can be mastered by the primary care practitioner and can lead to improved patient satisfaction. These techniques can be practiced using pig's feet. These minor plastic surgery techniques are an important component in a primary care practice.


Shape from recognition: a novel approach for 3-D face shape recovery.
In this paper, we develop a novel framework for robust recovery of three-dimensional (3-D) surfaces of faces from single images. The underlying principle is shape from recognition, i.e., the idea that pre-recognizing face parts can constrain the space of possible solutions to the image irradiance equation, thus allowing robust recovery of the 3-D structure of a specific part. Parts of faces like nose, lips and eyes are recognized and localized using robust expansion matching filter templates under varying pose and illumination. Specialized backpropagation based neural networks are then employed to recover the 3-D shape of particular face parts. Representation using principal components allows to efficiently encode classes of objects such as nose, lips, etc. The specialized networks are designed and trained to map the principal component coefficients of the part images to another set of principal component coefficients that represent the corresponding 3-D surface shapes. To achieve robustness to viewing conditions, the network is trained with a wide range of illumination and viewing directions. A method for merging recovered 3-D surface regions by minimizing the sum squared error in overlapping areas is also derived. Quantitative analysis of the reconstruction of the surface parts in varying illumination and pose show relatively small errors, indicating that the method is robust and accurate. Several examples showing recovery of the complete face also illustrate the efficacy of the approach.


Quantifying Spontaneous Ca2+ Fluxes and their Downstream Effects in Primary Mouse Midbrain Neurons.
Parkinson's disease (PD) is a devastating neurodegenerative disorder caused by the degeneration of dopaminergic (DA) neurons. Excessive Ca2+ influx due to the abnormal activation of glutamate receptors results in DA excitotoxicity and has been identified as an important mechanism for DA neuron loss. In this study, we isolate, dissociate, and culture midbrain neurons from the mouse ventral mesencephalon (VM) of ED14 mouse embryos. We then infect the long-term primary mouse midbrain cultures with an adeno-associated virus (AAV) expressing a genetically encoded calcium indicator, GCaMP6f under control of the human neuron-specific synapsin promoter, hSyn. Using live confocal imaging, we show that cultured mouse midbrain neurons display spontaneous Ca2+ fluxes detected by AAV-hSyn-GCaMP6f. Bath application of glutamate to midbrain cultures causes abnormal elevations in intracellular Ca2+ within neurons and this is accompanied by caspase-3 activation in DA neurons, as demonstrated by immunostaining. The techniques to identify glutamate-mediated apoptosis in primary mouse DA neurons have important applications for the high content screening of drugs that preserve DA neuron health.


Expression of cysteine protease genes in pea nodule development and senescence.
Coding sequences for two cysteine proteases were amplified from cDNA derived from pea nodule mRNA using primers based on conserved regions of plant cysteine proteases. One of the amplified cDNA sequences corresponded to a previously described cysteine protease gene, Cyp15a, expressed in pea shoots in response to dehydration (J.T. Jones and J.E. Mullet, Plant Mol. Biol. 28:1055-1065, 1995). Inside the pea root nodule, in situ hybridization revealed that this gene is expressed strongly in the apical region and more weakly in the uninfected cortex and in the central infected tissue where nitrogen fixation takes place. The complete sequence of the cDNA corresponding to the other gene, PsCyp1, was obtained. Expression of this gene, which was studied both on RNA blots and in situ, showed good correlation with the onset of nodule senescence. In situ hybridization studies revealed that PsCyp1 was expressed in senescent infected tissue at the base of the nodule. This signal was just detectable in normal symbiotically wild-type nodules but was much stronger in the early senescing nodules formed by a symbiotically defective mutant of Rhizobium leguminosarum.


Intramuscular hepatitis B immune globulin combined with lamivudine for prophylaxis against hepatitis B recurrence after liver transplantation.
Immunoprophylaxis using intravenous (IV) hepatitis B immune globulin (HBIG) decreases the recurrence of hepatitis B virus (HBV) infection after orthotopic liver transplantation (OLT). However, IV HBIG is expensive, has significant side effects, and is inconvenient to administer. An alternative approach for prophylaxis using intramuscular (IM) HBIG and oral lamivudine was prospectively evaluated in this study. Ten consecutive patients with cirrhosis with HBV infection who underwent OLT were included in this study. Nine of 10 patients received lamivudine, 150 mg/d, for an average duration of 8.6 months before OLT. Two of 10 patients with detectable HBV DNA at the time of OLT received 10,000 U (45 mL) of IV HBIG daily for 7 consecutive days, followed by 5 mL of IM HBIG weekly for the next 3 weeks, then every 3 weeks. The other 8 patients were HBV DNA negative at OLT and received one dose of IV HBIG (45 mL) during surgery, followed by 5 mL of IM HBIG weekly for 4 weeks, then every 3 weeks. All patients received lamivudine, 150 mg/d, after OLT. During a mean follow-up of 15.6 months, 9 of 10 patients achieved a protective hepatitis B surface antibody (HBsAb) titer greater than 200 IU/L and had no evidence of HBV recurrence. One patient failed to develop an adequate HBsAb titer and developed histological and virological evidence of recurrence. One patient died unrelated to HBV recurrence. Our preliminary data suggest that this combination prophylaxis with IM HBIG and lamivudine is effective and potentially cost saving.