Unique Pattern of Protein-Bound Maillard Reaction Products in Manuka (Leptospermum scoparium) Honey.
As a unique feature, honey from the New Zealand manuka tree (Leptospermum scoparium) contains substantial amounts of dihydroxyacetone (DHA) and methylglyoxal (MGO). Although MGO is a reactive intermediate in the Maillard reaction, very little is known about reactions of MGO with honey proteins. We hypothesized that the abundance of MGO should result in a particular pattern of protein-bound Maillard reaction products (MRPs) in manuka honey. A protein-rich high-molecular-weight fraction was isolated from 12 manuka and 8 non-manuka honeys and hydrolyzed enzymatically. By HPLC-MS/MS, 8 MRPs, namely, N-ε-fructosyllysine, N-ε-maltulosyllysine, carboxymethyllysine, carboxyethyllysine (CEL), pyrraline, formyline, maltosine, and methylglyoxal-derived hydroimidazolone 1 (MG-H1), were quantitated. Compared to non-manuka honeys, the manuka honeys were characterized by high concentrations of CEL and MG-H1, whereas the formation of N-ε-fructosyllysine was suppressed, indicating concurrence reactions of glucose and MGO at the ε-amino group of protein-bound lysine. Up to 31% of the lysine and 8% of the arginine residues, respectively, in the manuka honey protein can be modified to CEL and MG-H1, respectively. CEL and MG-H1 concentrations correlated strongly with the MGO concentration of the honeys. Manuka honey possesses a special pattern of protein-bound MRPs, which might be used to prove the reliability of labeled MGO levels in honeys and possibly enable the detection of fraudulent MGO or DHA addition to honey.


Creating and sustaining a digital community of practice for quality improvement in South-East Asia during the COVID-19 pandemic.
Ensuring quality of care in Low and Middle Income countries (LMICs) is challenging. Despite the implementation of various quality improvement (QI) initiatives in public and private sectors, the sustenance of improvements continues to be a major challenge. A team of healthcare professionals in India developed a digital community of practice (dCoP) focusing on QI which now has global footprints. The dCoP was conceptualised as a multitiered structure and is operational online at www.nqocncop.org from August 2020 onwards. The platform hosts various activities related to the quality of care, including the development of new products, and involves different cadres of healthcare professionals from primary to tertiary care settings. The platform uses tracking indicators, including the cost of sustaining the dCoP to monitor the performance of the dCoP. Since its launch in 2020, dCoP has conducted over 130 activities using 13 tools with 25 940 registration and 13 681 participants. From April 2021, it has expanded to countries across the South-East Asia region and currently has participants from 53 countries across five continents. It has developed 20 products in four thematic areas for a targeted audience. dCoP is supporting mentoring of healthcare professionals from five countries in the South-East Asia region in their improvement journey. Acquiring new knowledge and improvement in their daily clinical practice has been reported by 93% and 80% of participants, respectively. The dCoP and its partners have facilitated the publication of nearly 40 articles in international journals. This dCoP platform has become a repository of knowledge for healthcare professionals in the South-East Asia region. The current paper summarises the journey of this innovative dCoP in an LMIC setting for a wider global audience.


The impact of extended adjuvant temozolomide in newly diagnosed glioblastoma multiforme: a meta-analysis and systematic review.
Surgical resection followed by concurrent radiation therapy and temozolomide (TMZ) chemotherapy is the current standard treatment for glioblastoma multiforme (GBM). The present metaanalysis investigated the impact of prolonged TMZ maintenance therapy (more than 6 cycles) in comparison with standard TMZ maintenance therapy (exactly six cycles) on overall survival (OS) and progression-free survival (PFS) of patients with GBM. A meta-analysis of the literature was conducted using Medline, PubMed, EMBASE and the Cochrane Library in accordance with PRISMA guidelines. Seven articles involving 1018 patients were included. The overall survival was higher in the case group (>6 cycles TMZ) compared to the control group (6 cycles TMZ) (Z=2.375, P=0.018). The lower and upper limits were between 1.002-10.467 months. The case group had higher progression-free survival compared with the control group (Z=3.84; P<0.001). The lower and upper limits were between 2.559-7.894 months. Evidence from this meta-analysis suggests that prolonged TMZ therapy compared to the standard 6-cycle TMZ therapy was associated with higher survival in patients with glioblastoma.


Heat shock protein 90 is required for increased DNA binding activity of activator protein-1, a heterodimer of Fos/JunD, in rheumatoid synovial cells under inflammatory stimuli.
We have studied the DNA binding profiles of activator protein-1 (AP-1) involved in synovial overgrowth and osteoporosis in rheumatoid arthritis (RA) in relation to the molecular chaperon heat shock protein 90 (HSP90). The AP-1 binding activity of the nuclear extracts of rheumatoid synovial cells was basically increased as compared with osteoarthritic synovial cells. Upon stimulation with inflammatory cytokines IL-1beta or TNFalpha, the AP-1 binding activity was further increased in rheumatoid synovial cells, and increased AP-1 protein was composed as heterodimers of Fos and JunD which was not known before as a major component of AP-1 in rheumatoid synovial cells. The increase of AP-1 binding activity as induced by inflammatory cytokines was specifically inhibited by geldanamycin, radicicol or herbimycin A, specific inhibitors of HSP90, while AP-1 protein was not decreased by geldanamycin. Further, HSP90 protein was not decreased by the inhibitors. The findings indicate that HSP90 is required for increased AP-1 binding activity of rheumatoid synovial cells under inflammatory stimuli and that AP-1 binding activity is inhibited by functionally inactivating HSP90 with the inhibitors.


Primary infertility in women exposed to diethylstilboestrol in utero.
In a selected group of 40 women who had been exposed to diethylstilboestrol in utero, 18 conceived without difficulty and 22 had primary infertility. Among those with primary infertility there was a significantly higher rate of anatomical structural defects and a greater tendency for menstrual disorders than in those without infertility. Thirteen (59%) of the women with primary infertility conceived, most after treatment with ovulation stimulating drugs. Spontaneous abortion and tubal pregnancy were frequent (47% and 10% respectively) and similar in both fertility groups. Of 13 infertile women examined, 4 (31%) had mild hyperprolactinemia--a hithero unreported finding for such women.


An evaluation of the preoperative hemoglobin level as a prognostic factor for oral squamous cell carcinoma.
Hypoxia seems to be an influencing factor for oral squamous cell carcinomas (SCC), and several immunohistochemical markers have been discussed in this regard. The aim of the present study was to evaluate preoperative hemoglobin levels as a prognostic factor for oral SCC. The files of 287 patients who had been treated for oral SCC between 1999 and 2008 were studied retrospectively. Hemoglobin levels between 1 and 5 days prior to surgical treatment were compared to Tumor (T)- and Nodal (N)- status, local recurrence, and lymph node metastases rate. The minimum follow-up period was 12 months. From a total of 287 patients with oral SCC, 205 (71.4%) were in the normal hemoglobin (Hb) group (female Hb ≥ 12.0 g/dl; male Hb ≥ 13.0 g/dl), 53 (18.5%) in the mild anemia (female Hb = 11.0-11.9 g/dl; male Hb = 11.0-12.9 g/dl), and 29 (10.1%) in the severe anemia group (female & male Hb<11.0 g/dl). Anemia was significant for the development of lymph node metastasis (p = 0.005) as well as for local recurrence (p = 0.001). No significant correlation was found to the initial T status (p = 0.183). Our data suggests that an Hb of below 11 g/dl contributes to and is an indicator for a poor prognosis. Consequently, pre-treatment Hb corrections may significantly improve outcome, but further investigations, including blood transfusion/application of erythropoietin due to tumor anemia, independent of intraoperative blood-loss are necessary to ascertain their role in an improved survival.


Platysmaplasty: Is it Possible to Pull the Platysma Effectively in the Medial or Lateral Direction?
The aim of this study was to investigate whether it is possible to pull the platysma effectively in the medial or lateral direction (medial platysmaplasty and lateral platysmaplasty) and to explore the anatomical basis of those findings.Six hemifaces from 3 fresh cadavers were dissected. After skin removal, the platysma was pulled upward and in the medial or lateral direction with the ulnar side of the palm. Its mobility was checked. In 2 volunteers, using wooden bar, the skin overlying platysma was pulled in superomedial and superolateral direction.The platysma ran diagonally from the acromio-deltoid region to the perioral and submental area. In all hemifaces, the platysma was attached to the mandible along its course. The platysma inserted into the mandibular body. At its medial portion (approximately halfway medially from the mentum to the angle; 4-5 cm), the attachment was so firm that it could not be moved horizontally. The posterolateral portion of the platysma was indirectly attached to the mandible and movable. In cadaver, platysma did not move much when it was pulled in the medial direction. In the lateral direction, however, platysma did move well. In living body, when skin overlying platysma was pulled in superomedial direction and superolateral direction, 3 points marked on mandibular border moved about 1.5-2.0 cm and 2.0-2.5 cm respectively.It is thought that medial platysmaplasty can correct anterior neck deformities and redistribute neck skin mainly in the submental area, while that lateral platysmaplasty can pull the cheek skin in superolateral direction.


Quality evaluation of different honey samples produced in Peshawar valley.
The present study was designed to evaluate different honey samples obtained from local market for their quality parameters for assessment of their feasibility for foreign export by comparing it with international standards. The study was conducted at PCSIR laboratories complex, Peshawar, during 2006. The tested samples were evaluated for moisture content, Ash percentage, acid content, HMF and reducing sugars percentage. The moisture content of locally produced honey was in the range of 14.5 to 18.23%. The ash content of locally produced honey samples ranged between 0.047-0.35 which is within the standard limits. The acid content of the honey samples ranged between 19.5 and 38.0 meq kg(-1). The HMF contents of locally produced honeys ranged from 5.3 to 23.20 mg kg(-1). The content of reducing sugar of the tested samples ranged between 43.14 and 81.40% for the tested samples of locally produced honey. All of the samples were found to be in acceptable range of international standards for all of the tested parameters except for only one sample with lower reducing sugars. These samples were marked to be according to the international standards and are healthy for human consumption.


Immune responses to myxoma virus.
Myxoma virus causes the systemic disease myxomatosis in the European rabbit (Oryctolagus cuniculus). Originating in the South American rabbit Sylvilagus brasiliensis, where it causes a relatively localized fibroma, myxoma virus is a classic example of a virus that has jumped species to produce an exotic disease and then coevolved with its new host. Like other poxviruses, myxoma virus encodes multiple proteins capable of downregulating the host innate and acquired immune responses. Other virus-encoded proteins enable replication in host lymphocytes and monocytes, for example, by inhibiting apoptosis. Detailed studies on these proteins have demonstrated novel methods of interactions with the host immune system and added substantially to the understanding of the interaction of large DNA viruses with their hosts. Despite the increasingly detailed molecular knowledge of myxoma virus, relatively little is known about the dynamics of the interaction of the virus with the integrated host-immune system during infection and, in particular, about the evolution of resistance to the virus in wild rabbits or the species barrier. This review will focus on the detailed molecular studies that have been done with myxoma virus and discuss the more limited knowledge of the pathogenesis of myxoma virus in rabbits and the ways that the consolidated immune responses may determine genetic resistance to myxomatosis.


The use of a screening device to assess psychopathy in young offenders.
The aim of this research was to determine to what extent a psychopath screening device (the APSD) is useful in forensic assessments to predict general and violent offending. For this purpose, a cross-sectional study was done and 238 young people serving a sentence were assessed. The gold standard instrument used to measure psychopathy was the Psychopathy Checklist: Youth Version (PCL:YV; Forth, Kosson & Hare, 2003). The results indicate that the association found between the screening device scores and several indicators of risk is low if compared with those obtained with the PCL:YV, suggesting that it is less useful as a tool in order to predict offending or violent offences. However, an Area Under the Curve of .784 and a validity index of 62.5 support its use as a screening device or as a preliminary approach to assess psychopathy in this population. The usefulness of this instrument to make assessments with young people in the forensic setting is discussed.


64-slice computed tomography of bovine internal mammary artery coronary grafts.
In cases where conventional aortocoronary grafts cannot be used, No-React bovine internal mammary artery is a possible alternative. The aim of this study was to assess the patency and clinical performance of bovine internal mammary artery as a coronary bypass conduit, using 64-slice computed tomography coronary angiography. Eleven patients (mean age, 68.2 + or - 5.9 years) received 11 bovine grafts between 2002 and 2006. Eight of these patients were alive after a mean follow-up of 29.4 + or - 16.3 months. Their mean additive EuroSCORE was 6.5 + or - 3.2. The mean number of distal anastomoses was 2.5 + or - 0.5. Six grafts were anastomosed to the right coronary artery, 2 to the left anterior descending artery, and 3 to the circumflex artery. All 11 bovine grafts were found to be occluded after 14.1 + or - 3.6 months. This demonstrates very poor results with the bovine coronary graft, thus we do not recommend its use, and we suggest considering an hybrid approach in selected cases.


[Effect of the procedure of blood exchange on several indices of skeletal muscle metabolism].
In 85% blood exchange in healthy dogs and also in the animals under conditions of hemotransfusion shock a study was made of the nitrogen fraction contents, the activity of aspartic and alanine transpherases in the skeletal muscle in the course of 7 days. Blood exchange in hemotransfusion shock was highly effective. However, the process of "washing out" of the non-protein substances from the tissue in these animals was considerably lowered in comparison with that in healthy animals.


The effect of breast reconstruction surgery on body image among women after mastectomy: a meta-analysis.
The purpose of this study was to analyze the available evidence concerning the effects of breast reconstruction (BR) on body image of women with breast cancer. BR is a potential intervention to improve the body image of women with breast cancer. Conflicting research findings about the effects of breast reconstructive surgery on body image compared to breast conservative surgery (BCS) or mastectomy exist. A meta-analysis of studies found in MEDLINE, CINAHL, PsychINFO, PubMed as well as PQDT (dissertation and theses), and other unpublished literature resources from 1970 to 2010 were searched. Comparable studies that examined the difference of body image between breast cancer women receiving mastectomy plus BR and those with BCS were included. A clear definition about the domains of body image including body stigma, body concern, and transparency were identified. Mean effect sizes were calculated. Twelve studies were included in the meta-analysis. Women with BR and BCS were no different on the body concern domain of body image. However, women with BR had a significantly worse score on body stigma domain of body image than women receiving BCS. Women with BR had a better body image score than women with mastectomy. Women who are satisfied with their body shape may still perceive deficiencies because of the stigma of mastectomy and affect on body image. Consistent application of valid and reliable measures of body image specific to breast cancer women is needed.


The incidence of AIDS-defining illnesses in 4883 patients with human immunodeficiency virus infection. Royal Free/Chelsea and Westminster Hospitals Collaborative Group.
Acquired immunodeficiency syndrome (AIDS)-defining illnesses are known to occur at different levels of immunosuppression, and the incidence of diagnoses may also vary according to the CD4 lymphocyte count strata. Information about the incidence of disease at different levels of immunosuppression would help clinicians monitoring patients and would allow prophylaxis to be targeted at the most appropriate population. Between 1982 and July 1995, 4883 patients testing positive for the human immunodeficiency virus were seen at either the Royal Free or Chelsea and Westminster Hospitals in London, England. The incidence of each diagnosis, both overall and stratified by CD4 lymphocyte count, was calculated using a person-years analysis. Patients who had no CD4 lymphocyte counts measured during follow-up were excluded from the analysis. During a median follow-up period of 27.6 months, 3875 AIDS-defining illnesses were reported in 1713 patients. The incidence of AIDS-defining illnesses ranged from 6.22 per 100 person-years of follow-up for Pneumocystis carinii pneumonia (95% confidence interval, 5.74-6.70) to 0.37 for extrapulmonary tuberculosis (95% confidence interval, 0.26-0.48). The incidence of each AIDS-defining illness increased as the CD4 lymphocyte count declined; diagnoses such as cytomegalovirus and Mycobacterium avium-intracellulare complex infection had a low incidence at CD4 lymphocyte counts above 0.05x10(9)/L (50/mm3), while Kaposi sarcoma, P carinii pneumonia, and esophageal candidiasis had a high incidence throughout all CD4 lymphocyte count strata. This study provides important information about the risk of AIDS-defining illnesses at lower CD4 lymphocyte counts, enabling disease-specific prophylaxis to be targeted at the most appropriate population. In the future, as more prophylactic therapies are developed, this study will provide historical data of the incidence of diseases before specific prophylaxis was introduced.


The requirement of tumour necrosis factor-alpha and interferon-gamma for the expression of protective immunity to secondary murine tularaemia depends on the size of the challenge inoculum.
The present study was conducted to determine the extent to which the cytokines tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) are required to protect against primary or secondary murine tularaemia caused by the live vaccine strain of the facultative intracellular bacterium Francisella tularensis. It is shown that non-immune mice treated with neutralizing monoclonal antibodies (mAbs) against TNF-alpha and IFN-gamma are rendered defenceless against otherwise sublethal intravenous inocula of the bacterium. Treatment with either of the anti-cytokine mAbs resulted in even a very small inoculum of 500 c.f.u. of the pathogen multiplying unrestrictedly in the livers, spleens and lungs of non-immune mice to rapidly reach lethal numbers. By contrast, Francisella-immune mice treated with either of the mAbs remained capable of resolving secondary infection with 50-fold larger inocula. However, the need for TNF-alpha and IFN-gamma for controlling secondary tularaemia became critical when challenge inocula exceeded 10(6) c.f.u. Overall, the results imply that different defence mechanisms operate to control primary versus secondary murine tularaemia. Additionally, they show that the need for TNF-alpha and IFN-gamma to combat secondary infection depends on the size of the challenge inoculum.


Asymmetry and dynamics of a narrow sonar beam in an echolocating harbor porpoise.
A key component in the operation of a biosonar system is the radiation of sound energy from the sound producing head structures of toothed whales and microbats. The current view involves a fixed transmission aperture by which the beam width can only change via changes in the frequency of radiated clicks. To test that for a porpoise, echolocation clicks were recorded with high angular resolution using a 16 hydrophone array. The beam is narrower than previously reported (DI = 24 dB) and slightly dorso-ventrally compressed (horizontal -3 dB beam width: 13°, vertical -3 dB beam width: 11°). The narrow beam indicates that all smaller toothed whales investigated so far have surprisingly similar beam widths across taxa and habitats. Obtaining high directionality may thus be at least in part an evolutionary factor that led to high centroid frequencies in a group of smaller toothed whales emitting narrow band high frequency clicks. Despite the production of stereotyped narrow band high frequency clicks, changes in the directionality by a few degrees were observed, showing that porpoises can obtain changes in sound radiation.


Dual targeted poplar ferredoxin NADP(+) oxidoreductase interacts with hemoglobin 1.
Previous reports have connected non-symbiotic and truncated hemoglobins (Hbs) to metabolism of nitric oxide (NO), an important signalling molecule involved in wood formation. We have studied the capability of poplar (Populus tremula × tremuloides) Hbs PttHb1 and PttTrHb proteins alone or with a flavin-protein reductase to relieve NO cytotoxicity in living cells. Complementation tests in a Hb-deficient, NO-sensitive yeast (Saccharomyces cerevisiae) Δyhb1 mutant showed that neither PttHb1 nor PttTrHb alone protected cells against NO. To study the ability of Hbs to interact with a reductase, ferredoxin NADP(+) oxidoreductase PtthFNR was characterized by sequencing and proteomics. To date, by far the greatest number of the known dual-targeted plant proteins are directed to chloroplasts and mitochondria. We discovered a novel variant of hFNR that lacks the plastid presequence and resides in cytosol. The coexpression of PttHb1 and PtthFNR partially restored NO resistance of the yeast Δyhb1 mutant, whereas PttTrHb coexpressed with PtthFNR failed to rescue growth. YFP fusion proteins confirmed the interaction between PttHb1 and PtthFNR in plant cells. The structural modelling results indicate that PttHb1 and PtthFNR are able to interact as NO dioxygenase. This is the first report on dual targeting of central plant enzyme FNR to plastids and cytosol.


[Hemorheological aspects of the Arvin therapy of acute thromboses of the magistral veins of the lower extremities].
The rheologic blood properties were studied in patients with acute venous thrombosis during Arvin therapy. A marked correcting effect of the agent in removing the hemorheological disorders was demonstrated. It was found that the action of Arvin, linked with reduction of fibrinogen concentration, is attended by a decrease of blood viscosity, diminished aggregation of blood formed elements, and improvement of the functional condition of the microcirculatory bed during the whole course of treatment. The duration of the positive hemorheological effect of Arvin exceeds that in treatment by a complex of antithrombotic agents.


RX-3117 (Fluorocyclopentenyl-Cytosine)-Mediated Down-Regulation of DNA Methyltransferase 1 Leads to Protein Expression of Tumor-Suppressor Genes and Increased Functionality of the Proton-Coupled Folate Carrier.
(1) Background: RX-3117 (fluorocyclopentenyl-cytosine) is a cytidine analog that inhibits DNA methyltransferase 1 (DNMT1). We investigated the mechanism and potential of RX-3117 as a demethylating agent in several in vitro models. (2) Methods: we used western blotting to measure expression of several proteins known to be down-regulated by DNA methylation: O6-methylguanine-DNA methyltransferase (MGMT) and the tumor-suppressor genes, p16 and E-cadherin. Transport of methotrexate (MTX) mediated by the proton-coupled folate transporter (PCFT) was used as a functional assay. (3) Results: RX-3117 treatment decreased total DNA-cytosine-methylation in A549 non-small cell lung cancer (NSCLC) cells, and induced protein expression of MGMT, p16 and E-cadherin in A549 and SW1573 NSCLC cells. Leukemic CCRF-CEM cells and the MTX-resistant variant (CEM/MTX, with a deficient reduced folate carrier) have a very low expression of PCFT due to promoter hypermethylation. In CEM/MTX cells, pre-treatment with RX-3117 increased PCFT-mediated MTX uptake 8-fold, and in CEM cells 4-fold. With the reference hypomethylating agent 5-aza-2'-deoxycytidine similar values were obtained. RX-3117 also increased PCFT gene expression and PCFT protein. (4) Conclusion: RX-3117 down-regulates DNMT1, leading to hypomethylation of DNA. From the increased protein expression of tumor-suppressor genes and functional activation of PCFT, we concluded that RX-3117 might have induced hypomethylation of the promotor.


Analysis of HLA-E peptide-binding specificity and contact residues in bound peptide required for recognition by CD94/NKG2.
The MHC class Ib molecule HLA-E is the primary ligand for CD94/NKG2A-inhibitory receptors expressed on NK cells, and there is also evidence for TCR-mediated recognition of this molecule. HLA-E preferentially assembles with a homologous set of peptides derived from the leader sequence of class Ia molecules, but its capacity to bind and present other peptides remains to be fully explored. The peptide-binding motif of HLA-E was investigated by folding HLA-E in vitro in the presence of peptide libraries derived from a nonameric leader peptide sequence randomized at individual anchor positions. A high degree of selectivity was observed at four of five total anchor positions, with preference for amino acids present in HLA-E-binding peptides from class Ia leader sequences. Selectivity was also observed at the nonanchor P5 position, with preference for positively charged amino acids, suggesting that electrostatic interactions involving the P5 side chain may facilitate assembly of HLA-E peptide complexes. The observed HLA-E peptide-binding motif was strikingly similar to that previously identified for the murine class Ib molecule, Qa-1. Experiments with HLA-E tetramers bearing peptides substituted at nonanchor positions demonstrated that P5 and P8 are primary contact residues for interaction with CD94/NKG2 receptors. A conservative replacement of Arg for Lys at P5 completely abrogated binding to CD94/NKG2. Despite conservation of peptide-binding specificity in HLA-E and Qa-1, cross-species tetramer-staining experiments demonstrated that the interaction surfaces on CD94/NKG2 and the class Ib ligands have diverged between primates and rodents.


Desmoplastic small round cell tumor.
Desmoplastic small round cell tumor is an undifferentiated tumor associated with serosal surfaces, especially the peritoneum. It is found predominantly in adolescents and young adults and is much more common in males than in females. The tumor has a characteristic histology, with extensive stromal tissue around islands of small and undifferentiated cells revealing the desmoplastic appearance. The coexpression of epithelial and mesenchymal antigens distinguishes this entity from other small round and blue cell tumors of this age group. Cytogenetic studies showed a t(11;22) translocation that differs from the Ewing's tumor translocation and seems to be specific to this entity. The involvement of the WT1 and EWS genes in the translocation makes this tumor an interesting subject for research on histogenesis and differentiation in small round and blue cell tumors.


Novel high butanol production from lactic acid and pentose by Clostridium saccharoperbutylacetonicum.
We previously reported a butanol production process with pH-stat continuous feeding of dl-lactic acid and glucose as the co-substrate (Oshiro et al., Appl. Microbiol. Biotechnol., 87, 1177-1185, 2010). To accomplish butanol production from completely inedible substrates, in this study, we investigated acetone-butanol-ethanol (ABE) fermentation of Clostridium saccharoperbutylacetonicum N1-4 with lactic acid by using pentose as the co-substrate. Examination for optimum co-substrate indicated that arabinose was superior to glucose and xylose for ABE fermentation. Actually batch culture with lactic acid and arabinose without pH control exhibited higher butanol production (7.11 g/l) and lactic acid consumption (2.02 g) than those (6.62 g/l and 1.45 g, respectively) with glucose. Fed-batch culture without pH control increased these values to 12.08 g/l and 15.60 g/l butanol production, and to 3.83 g and 5.91 g lactic acid consumption by feeding the substrate once and twice, respectively. Finally, the result of gas chromatography-mass spectroscopy analysis using [1,2,3-(13)C(3)]-lactic acid indicated that lactic acid was converted to butanol with the efficiency of 51.9%. Thus, we established a novel high butanol production from lactic acid using arabinose as the co-substrate in simple fed-batch culture.


Association of autoantibodies to BP180 with disease activity in Greek patients with bullous pemphigoid.
39 bullous pemphigoid (BP) patients were studied to assess the clinical significance of anti-BP180 and anti-BP230 circulating autoantibodies of BP and correlate their titers with the clinical scores of the BP Disease Area Index (BPDAI) and the Autoimmune Bullous Skin Disorder Intensity Score (ABSIS) as well as with the intensity of pruritus measured by the BPDAI pruritus component. All parameters were evaluated by the time of diagnosis (baseline), month 3, and month 6. Titers of anti-BP180 autoantibodies were strongly correlated with BPDAI (r = 0.557, P value < 0.0001) and ABSIS (r = 0.570, P value < 0.0001) values, as well as with BPDAI component for the intensity of pruritus (rho = 0.530, P value = 0.001) at baseline. At month 3, titers of anti-BP180 autoantibodies were strongly correlated with BPDAI (rho = 0.626, P value = 0.000) and ABSIS (rho = 0.625, P value = 0.000) values, as well as with the BPDAI component for the intensity of pruritus (rho = 0.625, P value = 0.000). At month 6, titers of anti-BP180 autoantibodies were strongly correlated with BPDAI (rho = 0.527, P value = 0.001) and ABSIS (rho = 0.526, P value = 0.001) values, as well as with the BPDAI component for the intensity of pruritus (rho = 0.525, P value = 0.001). There was no statistically significant correlation between titers of anti-BP230 autoantibodies and the BPDAI, ABSIS, and BPDAI component for the intensity of pruritus at the same time points.


A FISH karyotype to study chromosome polymorphisms for the Elytrigia elongata E genome.
Elytrigia elongata (Host) Nevski(= Agropyron elongatum, Thinopyrum elongatum, 2n = 2x = 14, EE) has long been used as a source of various types of resistance for wheat improvement, and numerous transfers have been made. However, despite heavy use, no high-resolution karyotype exists. We characterized the E. elongata karyotype of several accessions applying highly repetitive DNA sequences as mcFISH probes for chromosome identification. The complete E. elongata disomic chromosome addition series and 11 ditelosomic addition lines in Chinese Spring wheat were exposed to sequential GISH-mcFISH. Based on the mcFISH results, each complete chromosome and each telocentric studied was unambiguously identified. The validation of the karyotype in 4 E. elongata accessions with different geographical origins showed extensive variations in the probe hybridization patterns, but this did not prevent chromosome identification. The established karyotype will be useful for the rapid identification of potential donor chromosomes in wheat improvement programs, allowing appropriate alien transfer.


Development and distribution of mast cells and neuropeptides in human fetus duodenum.
To study the developmental regularities and heterogeneity of mast cells (MC) in human fetus duodenum and the distribution and developmental regularities of substance P(SP), calcitonin gene-related peptide (CGRP)-immunoreactive (IR) peptidergic nerves in fetus duodenum, as well as the relationship between MC, SP and CGRP- IR peptidergic nerves. Duodena from 21 cases of human fetus and one term infant were stained by hematoxylin-eosin (HE), toluidine blue (TB) and immunohistochemical avidin-biotinylated peroxidase complex (ABC) method. Lobe-shape intestinal villi in duodenum were already developed at the twelfth week. At the 21st wk, muscular mucosa appeared gradually, and four layers were observed in the wall of duodenum. TB staining showed that the granules in the immature MC were pale violet, while the mature MC were strong violet in color by TB staining. Connective tissue MC (CTMC) appeared occasionally in submucosa and muscular layer of duodenum at the 16th wk. While the mucosa MC (MMC) appeared at the 18th wk. At the 22nd wk, both CTMC and MMC were activated, and distributed in the surrounding blood vessels and ganglions. The verge of some MC were unclear, and showed degranular phenomena. At the 14th wk, SP and CGRP-IR nerve fibers and cells appeared in the myenteric and submucous plexuses in small intestine, and the responses were turn strongly. Neurons were light to deep brown, and nerve fibers were present as varicose and liner profiles. On the corresponding site of serial sections, SP and CGRP immunohistochemical reactions were coexisted in one nerve fiber or cell. Some of MC showed SP and CGRP-IR positive staining. There are two heterogeneous kinds of MC in duodenum, MMC and CTMC. MC might play an important role in regulating blood circulation and sensation.


The role of hyperhomocysteinemia in neurological features associated with coeliac disease.
Although a range of neurological and psychiatric disorders are widely reported to be associated with coeliac patients, their pathogenesis remains unclear. Some such disorders are believed to be secondary to vitamin deficiency due to malabsorption, others to immune mechanisms. We hypothesise that hyperhomocysteinemia might, by damaging the blood-brain barrier, expose neuronal tissue to all neuro-irritative metabolites, such as homocysteine itself, a neurotoxic excitatory and proconvulsant amino acid. Neurons respond to these stimuli through hyperexcitability, thereby predisposing subjects to neurological disorders such as epilepsy and headache. Furthermore, persisting endothelial damage may cause blood extravasation and subsequent deposition of calcium salts. We suggest that this might be the pathogenesis of the CEC syndrome, which is characterized by the association of coeliac disease, epilepsy and cerebral calcifications. Indeed, homocysteine plays a well-known role in cardiovascular endothelial dysfunction, with high serum and cerebrospinal fluid levels often being reported in coeliac patients. Moreover, data in the literature show a strong, growing association of homocysteine with epilepsy and migraine in non-coeliac subjects. Despite these findings, homocysteine has never been held directly responsible for neuronal functional features (neuronal hyperexcitability underlying epilepsy and migraine) and structural brain damage (expressed as cerebral calcification) in coeliac patients. Damage to the blood-brain barrier might also facilitate immune reactions against neuronal tissue to a considerable extent. This hypothesis combines the two afore-mentioned theories (vitamin deficiency due to malabsorption and immune mechanisms). We also wish to point out that no studies have yet investigated the prevalence of neuronal hyperexcitability and subclinical electroencephalic abnormalities in children and adults with newly-diagnosed coeliac disease before the introduction of a gluten-free diet, and in particular any changes following the introduction of the diet. We believe that the onset of clinical symptoms such as migraine and convulsions is preceded by a period in which damage is expressed exclusively by subclinical electroencephalic abnormalities; persisting damage to neuronal tissue subsequently leads to clinical manifestations. We propose two types of investigations: the first is to determine whether newly-diagnosed coeliac patients with hyperhomocysteinemia are a subgroup at risk for neurological features (clinical and subclinical); the second is to determine whether appropriate treatment of hyperhomocysteinemia and vitamin B status deficiency improves neurological abnormalities and reduces the risk of cerebral calcifications. The aim of these investigations is to develop new therapeutic strategies designed to prevent neuronal damage and increase the quality of life in children affected by such disorders.


Relationships between nutrition screening checklists and the health and well-being of older Australian women.
To examine associations between nutrition screening checklists and the health of older women. Cross-sectional postal survey including measures of health and health service utilisation. as well as the Australian Nutrition Screening Initiative (ANSI), adapted from the Nutrition Screening Initiative (NSI). Australia, 1996. In total, 12,939 women aged 70-75 years randomly selected as part of the Australian Longitudinal Study on Women's Health. Responses to individual items in the ANSI checklist, and ANSI and NSI scores, were associated with measures of health and health service utilisation. Women with high ANSI and NSI scores had poorer physical and mental health, higher health care utilisation and were less likely to be in the acceptable weight range. The performance of an unweighted score (TSI) was also examined and showed similar results. Whereas ANSI classified 30% of the women as 'high-risk', only 13% and 12% were classified as 'high-risk' by the NSI and TSI, respectively. However, for identifying women with body mass index outside the acceptable range, sensitivity, specificity and positive predictive values for all of these checklists were less than 60%. Higher scores on both the ANSI and NSI are associated with poorer health. The simpler unweighted method of scoring the ANSI (TSI) showed better discrimination for the identification of 'at risk' women than the weighted ANSI method. The predictive value of individual items and the checklist scores need to be examined longitudinally.


Understanding the Giant Enhancement of Exchange Interaction in Bi_{2}Se_{3}-EuS Heterostructures.
A recent experiment indicated that a ferromagnetic EuS film in contact with a topological insulator Bi_{2}Se_{3} might show a largely enhanced Curie temperature and perpendicular magnetic anisotropy [F. Katmis et al., Nature (London) 533, 513 (2016).NATUAS0028-083610.1038/nature17635]. Through systematic density functional calculations, we demonstrate that in addition to the factor that Bi_{2}Se_{3} has a strong spin orbit coupling, the topological surface states are crucial to make these unusual behaviors robust as they hybridize with EuS states and extend rather far into the magnetic layers. The magnetic moments of Eu atoms are nevertheless not much enhanced, unlike what was reported in the experiment. Our results and model analyses provide useful insights for how these quantities are linked, and pave a way for the control of properties of magnetic films via contact with topological insulators.


In vivo "photofootprint" changes at sequences between the yeast GAL1 upstream activating sequence and "TATA" element require activated GAL4 protein but not a functional TATA element.
Transcription of the yeast GAL1 and GAL10 genes is induced by growth on galactose. Using the technique of photofootprinting in vivo, we previously documented equivalent transcription-dependent footprints within the putative "TATA" elements of both genes. To explore the functional significance of these observations, we created a 3-base-pair substitution mutation within the GAL1 promoter TATA element, which disrupted the ATATAA consensus sequence but left intact the photomodification targets. The mutation reduced galactose-induced RNA levels by a factor of 100. The mutant promoter no longer displayed the characteristic TATA sequence footprint, supporting the hypothesis that transcription activation involves the binding of a TATA box factor. We also observed a collection of transcription-correlated alterations in the modification pattern at sites between the UASG and the GAL1 TATA element, within sequences that are not required for inducible transcription. These patterns, characteristic of the induced wild-type GAL1 gene, were still galactose inducible with the TATA mutant GAl1 promoter, despite the low level of transcription from this promoter. We conclude that the GAL4-dependent protein/DNA structure responsible for the altered pattern within nonessential sequences is therefore not strictly coupled to an active TATA element or to high levels of expression. Nonetheless, the patterns probably reflect a stable protein-dependent structure that accompanies assembly of the transcription initiation complex.


Betanodavirus infection in the freshwater model fish medaka (Oryzias latipes).
Betanodaviruses, the causal agents of viral nervous necrosis in marine fish, have bipartite, positive-sense RNA genomes. As their genomes are the smallest and simplest among viruses, betanodaviruses have been studied in detail as model viruses by using a genetic-engineering system, as has occurred with the insect alphanodaviruses, the other members of the family Nodaviridae. However, studies of virus-host interactions have been limited, as betanodaviruses basically infect marine fish at early developmental stages (larval and juvenile). These fish are only available for a few months of the year and are not suitable for the construction of a reverse-genetics system. To overcome these problems, several freshwater fish species were tested for their susceptibility to betanodaviruses. It was found that adult medaka (Oryzias latipes), a well-known model fish, was susceptible to both Striped jack nervous necrosis virus (the type species of the genus Betanodavirus) and Redspotted grouper nervous necrosis virus (RGNNV), which have different host specificities in marine fish species. Infected medaka exhibited erratic swimming and the viruses were localized specifically in the brain, spinal cord and retina of the infected fish, similar to the pattern of infection in naturally infected marine fish. Moreover, medaka were susceptible to RGNNV at the larval stage. This is the first report of a model virus-model host infection system in fish. This system should facilitate elucidation of the mechanisms underlying RNA virus infections in fish.


Embryonic zeta-globin chains in adults: a marker for alpha-thalassemia-1 haplotype due to a greater than 17.5-kb deletion.
Hemolysates from adults with the alpha-thalassemia-1 haplotype due to the greater than 17.5-kb deletion of both alpha-globin genes from the same chromosome were found to contain embryonic zeta (zeta)-globin chains (alpha-globin-like chains), as determined by a specific and sensitive radioimmunoassay and an electrophoretic technique. zeta-Globin chains were not present in hemolysates from adults with deletion of a single alpha-globin gene from one or both chromosomes. These results indicate that zeta-globin chains, which can be assayed by immunologic techniques, can serve as markers for the alpha-thalassemia-1 haplotype due to the greater than 17.5-kb deletion. The ability to detect zeta-globin chains may be useful in populations in which the gene frequency of the greater than 17.5-kb deletion is high, for screening couples at risk of having offspring with homozygous alpha-thalassemia.


Evaluation of Low-Loss Polymer Switches for Multinuclear MRI/S.
Implementation of multinuclear MRI/S as a diagnostic tool in clinical settings faces many challenges. One of those challenges is the development of highly sensitive multinuclear RF coils. Current multi-tuning techniques incorporate lossy components that impact the highest achievable SNR for at least one of the coil frequencies. As a result, optimization of multinuclear coil designs continues to be a priority for RF hardware engineers. To address this challenge, a new frequency switching technology that incorporates stimuli-responsive polymer materials was explored. Q measurements were used as a comparison metric between single-tuned, a standard switching network, and the proposed switching technology. The Q losses measured in the new switching method remained below 38% when compared to single-tuned coils. These results are consistent with low loss values reported using traditional switching networks. Furthermore, preliminary testing indicates that there is potential for improvement. These results establish the new technology as a promising alternative to traditional switching techniques.Clinical Relevance- A low loss multi-tuning technique for MRI radiofrequency coils has the potential of improving the study and diagnosis of disease.


Oral fluid as a medium for the detection of hepatitis B surface antigen.
Currently viral antigens and antibodies are detected by traditional serological tests. However, the introduction of oral fluid as an alternative medium would allow other alternatives. The collection of oral fluid is, in comparison with venepuncture, less invasive, less painful, less expensive (i.e., no trained personal required), and safe (prevention of needle stick injuries). Also large numbers of samples can be collected easily for epidemiological purposes. Forty-three HBsAg positive and seventy-three HBsAg negative paired serum/oral fluid samples were tested. They were collected from patients attending university hospitals. The oral fluid samples were collected using the Oracol collection device and they were subjected to an IgG quantification assay to ensure their quality and quantity. The detection of HBsAg in oral fluid was carried out using a modified ETI-MAK-4 ELISA. The validation of this oral fluid test gave a sensitivity and specificity of 90.7% and 100%, respectively. The modified ETI-MAK-4 ELISA is a suitable test for oral fluid samples collected by the Oracol collection device for epidemiological purposes.


[Objective assessment of the severity of gunshot wounds].
By the present time, many mathematical models have been developed for assessing the status of patients with grave mechanical injuries. However, comparative analysis of the efficacy of predicting the outcomes of shock and traumatic disease with the use of some models in patients with war and other than war traumas showed that these methods are not fit for cases with gunshot wounds. We consider that a differentiated approach is needed in assessment of the status of wounded and injured patients.


Dust and Cobalt Levels in the Austrian Tungsten Industry: Workplace and Human Biomonitoring Data.
In general, routine industrial hygiene (IH) data are collected not to serve for scientific research but to check for compliance with occupational limit values. In the preparation of an occupational retrospective cohort study it is vital to test the validity of the exposure assessment based on incomplete (temporal coverage, departments) IH data. Existing IH data from a large hard metal plant was collected. Individual workers' exposure per year and department was estimated based on linear regression of log-transformed exposure data for dust, tungsten, and cobalt. Estimated data were back-transformed, and for cobalt the validity of the estimates was confirmed by comparison with individual cobalt concentrations in urine. Air monitoring data were available from 1985 to 2012 and urine tests from the years 2008 to 2014. A declining trend and significant differences among departments was evident for all three air pollutants. The estimated time trend fitted the time trend in urine values well. At 1 mg/m³, cobalt in the air leads to an excretion of approximately 200 µg/L cobalt in urine. Cobalt levels in urine were significantly higher in smokers with an interaction effect between smoking and air concentrations. Exposure estimates of individual workers are generally feasible in the examined plant, although some departments are not documented sufficiently enough. Additional information (expert knowledge) is needed to fill these gaps.


The Biodiversity and Geochemistry of Cryoconite Holes in Queen Maud Land, East Antarctica.
Cryoconite holes are oases of microbial diversity on ice surfaces. In contrast to the Arctic, where during the summer most cryoconite holes are 'open', in Continental Antarctica they are most often 'lidded' or completely frozen year-round. Thus, they represent ideal systems for the study of microbial community assemblies as well as carbon accumulation, since individual cryoconite holes can be isolated from external inputs for years. Here, we use high-throughput sequencing of the 16S and 18S rRNA genes to describe the bacterial and eukaryotic community compositions in cryoconite holes and surrounding lake, snow, soil and rock samples in Queen Maud Land. We cross correlate our findings with a broad range of geochemical data including for the first time 13C and 14C analyses of Antarctic cryoconites. We show that the geographic location has a larger effect on the distribution of the bacterial community compared to the eukaryotic community. Cryoconite holes are distinct from the local soils in both 13C and 14C and their isotopic composition is different from similar samples from the Arctic. Carbon contents were generally low (≤0.2%) and older (6-10 ky) than the surrounding soils, suggesting that the cryoconite holes are much more isolated from the atmosphere than the soils.


Gold Nanoparticles: Promising Agent to Improve the Diagnosis and Therapy of Cancer.
Gold nanoparticles have been exploited for nanobiotechnology applications for the last two decades. New insights of the nanomaterials as promising agent for cancer diagnosis and therapy have just started to emerge. Due to the size- and shape-dependent optical, electrical and thermal properties, gold nanoparticles are being developed as diagnostic reagents, drug carriers, contrast agents, photothermal agents and radiosensitisers. This review aims to summarize the latest advances of gold nanoparticles in cancer treatment. We undertook a systematical search for research literatures using a well-framed review question and presented the applications in different fields, including early cancer diagnosis, imaging, radiotherapy, chemotherapy, gene therapy and photothermal therapy, which were fully described, filtered, combined and analyzed in order to provide documented proofs on the applications of gold nanoparticles in current cancer treatments. One hundred and sixty papers were included in the review, the majority of which represent latest researches in the field of gold nanoparticle-based diagnosis and therapy for cancer. Conventional treatment strategies for cancer cannot identify normal and cancer cells. While due to the high surface area to volume ratio and rich surface functionalization chemistry, gold nanoparticle can greatly enhance the targeting with adverse side effects of traditional treatment on normal tissues being avoided. Gold nanoparticles have greatly improved the traditional treatment due to their unique properties. However, their size-dependent toxicity, distribution and clearance need further studies to make them a clinical reality.


Intrinsic osteoinductivity of PCL-DA/PLLA semi-IPN shape memory polymer scaffolds.
Engineering osteoinductive, self-fitting scaffolds offers a potential treatment modality to repair irregularly shaped craniomaxillofacial bone defects. Recently, we innovated on osteoinductive poly(ε-caprolactone)-diacrylate (PCL-DA) shape memory polymers (SMPs) to incorporate poly-L-lactic acid (PLLA) into the PCL-DA network, forming a semi-interpenetrating network (semi-IPN). Scaffolds formed from these PCL-DA/PLLA semi-IPNs display stiffnesses within the range of trabecular bone and accelerated degradation relative to scaffolds formed from slowly degrading PCL-DA SMPs. Herein, we demonstrate for the first time that PCL-DA/PLLA semi-IPN SMP scaffolds show increased intrinsic osteoinductivity relative to PCL-DA. We also confirm that application of a bioinspired polydopamine (PD) coating further improves the osteoinductive capacity of these PCL-DA/PLLA semi-IPN SMPs. In the absence of osteogenic supplements, protein level assessment of human mesenchymal stem cells (h-MSCs) cultured in PCL-DA/PLLA scaffolds revealed an increase in expression of osteogenic markers osterix, bone morphogenetic protein-4 (BMP-4), and collagen 1 alpha 1 (COL1A1), relative to PCL-DA scaffolds and osteogenic medium controls. Likewise, the expression of runt-related transcription factor 2 (RUNX2) and BMP-4 was elevated in the presence of PD-coating. In contrast, the chondrogenic and adipogenic responses associated with the scaffolds matched or were reduced relative to osteogenic medium controls, indicating that the scaffolds display intrinsic osteoinductivity.


Repetitive transcranial magnetic stimulation for the treatment of obsessive compulsive disorder: a double-blind controlled investigation.
To determine the efficacy and tolerability of repetitive transcranial magnetic stimulation (rTMS) as a treatment for obsessive compulsive disorder (OCD) in a double-blind placebo-controlled study. Subjects with treatment-resistant OCD were randomized to rTMS (n = 10) or sham rTMS (n = 8) for 10 sessions of daily stimulation over the left dorsolateral prefrontal cortex (DLPFC), with subjects and raters being blind to the treatment. Subjects were offered an open extension of up to 20 sessions of rTMS. The two groups did not differ on change in Yale-Brown Obsessive Compulsive Scale (YBOCS) or Maudsley Obsessive-Compulsive Inventory scores over 10 sessions, with or without correction for depression ratings. Over 20 sessions, there was a significant reduction in total YBOCS scores, but not after controlling for depression. rTMS over 20 sessions was well tolerated. Two weeks of rTMS over the left DLPFC is ineffective for treatment-resistant OCD.


Drosophila mitoferrin is essential for male fertility: evidence for a role of mitochondrial iron metabolism during spermatogenesis.
Mammals and Drosophila melanogaster share some striking similarities in spermatogenesis. Mitochondria in spermatids undergo dramatic morphological changes and syncytial spermatids are stripped from their cytoplasm and then individually wrapped by single membranes in an individualization process. In mammalian and fruit fly testis, components of the mitochondrial iron metabolism are expressed, but so far their function during spermatogenesis is unknown. Here we investigate the role of Drosophila mitoferrin (dmfrn), which is a mitochondrial carrier protein with an established role in the mitochondrial iron metabolism, during spermatogenesis. We found that P-element insertions into the 5'-untranslated region of the dmfrn gene cause recessive male sterility, which was rescued by a fluorescently tagged transgenic dmfrn genomic construct (dmfrnvenus). Testes of mutant homozygous dmfrnSH115 flies were either small with unorganized content or contained some partially elongated spermatids, or testes were of normal size but lacked mature sperm. Testis squashes indicated that spermatid elongation was defective and electron micrographs showed mitochondrial defects in elongated spermatids and indicated failed individualization. Using a LacZ reporter and the dmfrnvenus transgene, we found that dmfrn expression in testes was highest in spermatids, coinciding with the stages that showed defects in the mutants. Dmfrn-venus protein accumulated in mitochondrial derivatives of spermatids, where it remained until most of it was stripped off during individualization and disposed of in waste bags. Male sterility in flies with the hypomorph alleles dmfrnBG00456 and dmfrnEY01302 over the deletion Df(3R)ED6277 was increased by dietary iron chelation and suppressed by iron supplementation of the food, while male sterility of dmfrnSH115/Df(3R)ED6277 flies was not affected by food iron levels. In this work, we show that mutations in the Drosophila mitoferrin gene result in male sterility caused by developmental defects. From the sensitivity of the hypomorph mutants to low food iron levels we conclude that mitochondrial iron is essential for spermatogenesis. This is the first time that a link between the mitochondrial iron metabolism and spermatogenesis has been shown. Furthermore, due to the similar expression patterns of some mitochondrial iron metabolism genes in Drosophila and mammals, it is likely that our results are applicable for mammals as well.


Biosynthesis of proteoglycan in vitro by cartilage from human osteochondrophytic spurs.
Proteoglycan biosynthesis by human osteochondrophytic spurs (osteophytes) obtained from osteoarthritic femoral heads at the time of surgical joint replacement was studied under defined culture conditions in vitro. Osteophytes were primarily present in two anatomic locations, marginal and epi-articular. Minced tissue slices were incubated in the presence of [(35)S]sulphate or [(14)C]glucosamine. Osteophytes incorporated both labelled precursors into proteoglycan, which was subsequently characterized by CsCl-isopycnic-density-gradient ultracentrifugation and chromatography on Sepharose CL-2B. The material extracted with 0.5m-guanidinium chloride showed 78.1% of [(35)S]sulphate in the A1 fraction after centrifugation. Only 23.0% of the [(35)S]sulphate in this A1 fraction was eluted in the void volume of Sepharose CL-2B under associative conditions. About 60-80% of the [(35)S]sulphate in the tissue 4m-guanidinium chloride extract was associated with monomeric proteoglycan (fraction D1). The average partition coefficient (K(av.)) of the proteoglycan monomer on Sepharose CL-2B was 0.28-0.33. Approx. 12.4% of this monomer formed stable aggregates with high-molecular-weight hyaluronic acid in vitro. Sepharose CL-2B chromatography of fractions with lower buoyant densities (fractions D2-D4) demonstrated elution profiles on Sepharose CL-2B substantially different than that of fraction D1, indicative of the polydisperse nature of the newly synthesized proteoglycan. Analysis of the composition and chain size of the glycosaminoglycans showed the following: (1) preferential elution of both [(35)S]sulphate and [(14)C]glucosamine in the 0.5m-LiCl fraction on DEAE-cellulose; (2) the predominant sulphated glycosaminoglycan was chondroitin 6-sulphate (60-70%), with 9-11% keratan sulphate in the monomer proteoglycan; (3) K(av.) values of 0.38 on Sephadex G-200 and 0.48 on Sepharose CL-6B were obtained with papain-digested and NaBH(4)-treated D1 monomer respectively. A comparison of the synthetic with endogenous glycosaminoglycans indicated similar types. These studies indicated that human osteophytes synthesized in vitro sulphated proteoglycans with some characteristics similar to those of mature human articular cartilage, notably in the size of their proteoglycan monomer and predominance of chondroitin 6-sulphate. They differed from articular cartilage primarily in the lack of substantial quantities of keratan sulphate and aggregation properties associated with monomer interaction with hyaluronic acid.


Spatial updating of environments described in texts.
People update egocentric spatial relations in an effortless and on-line manner when they move in the environment, but not when they only imagine themselves moving. In contrast to previous studies, the present experiments examined egocentric updating with spatial scenes that were encoded linguistically instead of perceived directly. Experiment 1 demonstrated that, regardless of the mode of rotation (physical or imagined), egocentric updating takes place in a deliberate and backward fashion when the locations of objects are anchored in a mental framework. Experiment 2 involved only imagined rotations and showed that results remained unchanged when spatial labels were removed from the scene descriptions. Experiment 3 provided evidence that physical rotations--but not imagined rotations--lead to on-line updating of egocentric relations, provided that the objects of the scene are represented in a sensorimotor framework. The present results suggest that physical movements and sensorimotor encoding are both prerequisites of effortless egocentric updating.


Calorie restriction does not elicit a robust extension of replicative lifespan in Saccharomyces cerevisiae.
Calorie restriction (CR) is often described as the most robust manner to extend lifespan in a large variety of organisms. Hence, considerable research effort is directed toward understanding the mechanisms underlying CR, especially in the yeast Saccharomyces cerevisiae. However, the effect of CR on lifespan has never been systematically reviewed in this organism. Here, we performed a meta-analysis of replicative lifespan (RLS) data published in more than 40 different papers. Our analysis revealed that there is significant variation in the reported RLS data, which appears to be mainly due to the low number of cells analyzed per experiment. Furthermore, we found that the RLS measured at 2% (wt/vol) glucose in CR experiments is partly biased toward shorter lifespans compared with identical lifespan measurements from other studies. Excluding the 2% (wt/vol) glucose experiments from CR experiments, we determined that the average RLS of the yeast strains BY4741 and BY4742 is 25.9 buds at 2% (wt/vol) glucose and 30.2 buds under CR conditions. RLS measurements with a microfluidic dissection platform produced identical RLS data at 2% (wt/vol) glucose. However, CR conditions did not induce lifespan extension. As we excluded obvious methodological differences, such as temperature and medium, as causes, we conclude that subtle method-specific factors are crucial to induce lifespan extension under CR conditions in S. cerevisiae.


Unique toroidal morphology from composition and sequence control of triblock copolymers.
The mechanism by which the unique toroidal supramolecular assemblies were formed for triblock copolymers of acrylic acid (AA), methyl acrylate (MA), and styrene (S), PAA99-b-PMA73-b-PS66, was probed in this study by investigating the influences of the block copolymer compositions and sequences. Two triblock copolymers, PAA99-b-PMA73-b-PS66 and PAA99-b-PS76-b-PMA62, and two diblock copolymers, PAA99-b-PMA155 and PAA99-b-PS133, were studied under experimental solution-state conditions that involved a range of solvent/nonsolvent (tetrahydrofuran/water) compositions, each in the presence of 2,2'-(ethylenedioxy)bis(ethylamine). The resulting morphologies were determined by transmission electron microscopy. The failures to afford toroidal supramolecular assemblies from both diblock copolymers having comparable lengths of the total hydrophobic chain segment, either entirely PMA or entirely PS, and from the triblock copolymer having a reversed connection sequence for the hydrophobic (PMA and PS) segments demonstrate the unique self-assembly behaviors of triblock copolymers and the importance of the block copolymer sequence.


Endorphins in septic shock: hemodynamic and endocrine effects of an opiate receptor antagonist and agonist.
The pathophysiological role of endorphins in septic shock was studied in a porcine model. Septic shock was induced by the intravenous infusion of live Escherichia coli. Naloxone hydrochloride, an opiate receptor blocker, given during profound septic shock, increased blood concentrations of glucagon and cyclic adenosine monophosphate (cAMP), while BP and cardiac output increased transiently. Heart rate and hepatic glycogen value decreased, but insulin and cortisol levels remained unchanged. In contrast, exogenous morphine injection produced further reduction of BP, increased pulmonary wedge pressure, and increased substance P, while growth hormone level and cardiac output remained unchanged. Neither hormonal nor hemodynamic changes were noted in saline controls. Thus, the endogenous opiates appear partly responsible for the hemodynamic derangements during septic shock, and naloxone is able to reverse such depression, even though the effects are transient and relatively minor when naloxone is given late in the course of septic shock. Endogenous opiates also affect the hormonal homeostasis in shock, and there are indications that this may be mediated by the adenylate cyclase-cAMP system.


Dioxin photoproducts of triclosan and its chlorinated derivatives in sediment cores.
Triclosan, a widely used antimicrobial, is known to undergo phototransformation in aqueous solution to form 2,8-dichlorodibenzo-p-dioxin (2,8-DCDD). Two sediment cores from a wastewater-impacted depositional zone of the Mississippi River were analyzed for triclosan by ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS-Q(3)) and for a suite of polychlorinated dioxins and furans by high resolution gas chromatography-mass spectrometry (HRGC-MS) to provide evidence of this photoreaction in the environment. 2,8-DCDD was detected at levels that trended with the historical use of triclosan since its introduction in the 1960s. Three other dioxin congeners, 2,3,7-TCDD, 1,2,8-TriCDD, and 1,2,3,8-TCDD, which are known photoproducts of chlorinated derivatives of triclosan, were also detected with similar trend profiles. These four congeners comprised the majority of di- through tetra-chlorinated dioxins. The trend profile of these specific dioxin congeners did not correlate with the trend profile of the higher-chlorinated dioxin homologues or any chlorinated furan homologues, suggesting a unique source. These results are fully consistent with the phototransformation of triclosan and its chlorinated derivatives that form during wastewater chlorine disinfection as the source of 2,8-DCDD, 2,3,7-TriCDD, 1,2,8-TriCDD, and 1,2,3,8-TCDD in this aquatic environment. As the levels of triclosan-derived dioxins increased over time and the total level of chlorinated dioxins decreased, the contribution of triclosan-derived dioxins to the total dioxin pool increased to as high as 31% by mass in recent years, indicating that their contribution to total dioxin toxicity may need consideration.


Catheter removal versus retention in the management of catheter-associated enterococcal bloodstream infections.
Enterococci are an important cause of central venous catheter (CVC)-associated bloodstream infections (CA-BSI). It is unclear whether CVC removal is necessary to successfully manage enterococcal CA-BSI. A 12-month retrospective cohort study of adults with enterococcal CA-BSI was conducted at a tertiary care hospital; clinical, microbiological and outcome data were collected. A total of 111 patients had an enterococcal CA-BSI. The median age was 58.2 years (range 21 to 94 years). There were 45 (40.5%) infections caused by Entercoccus faecalis (among which 10 [22%] were vancomycin resistant), 61 (55%) by Enterococcus faecium (57 [93%] vancomycin resistant) and five (4.5%) by other Enterococcus species. Patients were treated with linezolid (n=51 [46%]), vancomycin (n=37 [33%]), daptomycin (n=11 [10%]), ampicillin (n=2 [2%]) or quinupristin/dalfopristin (n=2 [2%]); seven (n=6%) patients did not receive adequate enterococcal treatment. Additionally, 24 (22%) patients received adjunctive gentamicin treatment. The CVC was retained in 29 (26.1%) patients. Patients with removed CVCs showed lower rates of in-hospital mortality (15 [18.3%] versus 11 [37.9]; P=0.03), but similar rates of recurrent bacteremia (nine [11.0%] versus two (7.0%); P=0.7) and a similar post-BSI length of hospital stay (median days [range]) (11.1 [1.7 to 63.1 days] versus 9.3 [1.9 to 31.8 days]; P=0.3). Catheter retention was an independent predictor of mortality (OR 3.34 [95% CI 1.21 to 9.26]). To the authors' knowledge, the present article describes the largest enterococcal CA-BSI series to date. Mortality was increased among patients who had their catheter retained. Additional prospective studies are necessary to determine the optimal management of enterococcal CA-BSI.


Supporting family carers in home-based end-of-life care: using participatory action research to develop a training programme for support workers and volunteers.
Family carers are crucial in enabling dying people to stay at home, but are often not prepared for their caring role, receiving little support from formal health and social care services. It is increasingly likely that any help or support family carers receive will be provided by a third sector organisation on either a voluntary basis or by untrained carer support workers. To produce a training programme designed to equip carer support workers and volunteers with the basic skills and knowledge needed to support family carers. Participatory action research, a collaborative form of working in which those who are affected by an issue take a lead role in the research, was used. Bereaved carers acting as research partners, support workers and representatives of third sector organisations took an active part in designing, developing, piloting and refining the programme in a number of interlinked stages. During development, the programme was piloted on four occasions and evaluated by 36 trainees and 3 trainers. The outcome of the project is an innovative, 1-day training programme, offering an introduction to supporting family carers who are looking after someone approaching the end of life. The use of participatory action research methods enabled the development of a programme that addresses support needs identified by bereaved carers and training needs identified by carer support workers.The finished programme includes all the materials necessary to run a training day for support workers and volunteers: facilitator's notes, trainee workbook, slides, promotional poster and pre-course reading for trainees. Knowledge of issues involved in end-of-life and palliative care is not required, although some experience in delivering training is advisable. The programme evaluated well during development, but further research is required to examine the transfer of learning into the workplace.


[Analysis of complications of operation for appendicular peritonitis in children].
Complications, occurred after the operation for appendicular peritonitis in children, were analyzed. The frequency of the complications occurrence in patients with an acute catarrhal appendicitis had constituted 1.4%, with phlegmonous one--3.5%, gangrenous--12.1%, gangrenous appendicitis with local restricted peritonitis--18.9%, with diffuse purulent peritonitis--42%. The dependence of the complications frequency on character and severity of primary changes in abdominal cavity was established.


International Consensus Recommendations for Safe Use of LAMS for On- and Off-label Indications Using a Modified Delphi Process.
The study aimed to develop international consensus recommendations on the safe use of lumen-apposing metal stents (LAMSs) for on- and off-label indications. Based on the available literature, statements were formulated and grouped into the following categories: general safety measures, peripancreatic fluid collections (PFC), biliary drainage (EUS-BD), gallbladder drainage (EUS-GBD), gastroenterostomy (EUS-GE) and gastric access temporary for endoscopy (GATE). The evidence level of each statement was determined using the GRADE methodology.International LAMS experts were invited to participate in a modified Delphi process. When no 80% consensus was reached, the statement was modified based on expert feedback. Statements were rejected if no consensus was reached after the third Delphi round. Fifty-six (93.3%) out of 60 formulated statements were accepted, of which 35 (58.3%) in the first round.Consensus was reached on the optimal learning path, preprocedural imaging, the need for airway protection and essential safety measures during the procedure, such as the use of Doppler and measurement of the distance between the gastrointestinal lumen and the target structure.Specific consensus recommendations were generated for the different LAMS indications, covering, among others, careful patient selection, the preferred size of the LAMS, the need for antibiotics, the preferred anatomic location of the LAMS, the need for coaxial pigtail placement, and the appropriate management of LAMS-related adverse events. Through a modified international Delphi process, we developed general and indication-specific experience- and evidence-based recommendations on the safe use of LAMS.


Polarized mitochondria as guardians of NK cell fitness.
Distinct metabolic demands accompany lymphocyte differentiation into short-lived effector and long-lived memory cells. How bioenergetics processes are structured in innate natural killer (NK) cells remains unclear. We demonstrate that circulating human CD56Dim (NKDim) cells have fused mitochondria and enhanced metabolism compared with CD56Br (NKBr) cells. Upon activation, these 2 subsets showed a dichotomous response, with further mitochondrial potentiation in NKBr cells vs paradoxical mitochondrial fission and depolarization in NKDim cells. The latter effect impaired interferon-γ production, but rescue was possible by inhibiting mitochondrial fragmentation, implicating mitochondrial polarization as a central regulator of NK cell function. NKDim cells are heterogeneous, and mitochondrial polarization was associated with enhanced survival and function in mature NKDim cells, including memory-like human cytomegalovirus-dependent CD57+NKG2C+ subsets. In contrast, patients with genetic defects in mitochondrial fusion had a deficiency in adaptive NK cells, which had poor survival in culture. These results support mitochondrial polarization as a central regulator of mature NK cell fitness.


Cloning and expression of Drosophila melanogaster UDP-GlcNAc:alpha-3-D-mannoside beta1,2-N-acetylglucosaminyltransferase I.
A TBLASTN search of the Drosophila melanogaster expressed sequence tag (EST) database with the amino acid sequence of human UDP-N-acetylglucosamine:alpha-3-D-mannoside beta-1,2-N-acetylglucosaminyltransferase I (GnT I, EC 18.104.22.168) as probe yielded a clone (GM01211) with 56% identity over 36 carboxy-terminal amino acids. A 550 base pair (bp) probe derived from the EST clone was used to screen a Drosophila cDNA library in lambda-ZAP II and two cDNAs lacking a start ATG codon were obtained. 5'-Rapid amplification of cDNA ends (5'-RACE) yielded a 2828 bp cDNA containing a full-length 1368 bp open reading frame encoding a 456 amino acid protein with putative N-terminal cytoplasmic (5 residues) and hydrophobic transmembrane (20 residues) domains. The protein showed 52% amino acid sequence identity to human GnT I. This cDNA, truncated to remove the N-terminal hydrophobic domain, was expressed in the baculovirus/Sf9 system as a secreted protein containing an N-terminal (His)6 tag. Protein purified by adsorption to and elution from nickel beads converted Man alpha1-6(Man alpha1-3)Man beta-octyl (M3-octyl) to Man alpha1-6(GlcNAc beta1-2Man alpha1-3)Man beta-octyl. The Km values (0.7 and 0.03 mM for M3-octyl and UDP-GlcNAc respectively), temperature optimum (37 degrees C), pH optimum (pH 5 to 6) and divalent cation requirements (Mn > Fe, Mg, Ni > Ba, Ca, Cd, Cu) were similar to mammalian GnT I. TBLASTN searches of the Berkeley Drosophila Genome Project database with the Drosophila GnT I cDNA sequence as probe allowed localization of the gene to chromosomal region 2R; 57A9. Comparison of the cDNA and genomic DNA sequences allowed the assignment of seven exons and six introns; all introns showed GT-AG splice site consensus sequences. This is the first insect GnT I gene to be cloned and expressed.


Relationship between the intracellular effects of camptothecin and the inhibition of DNA topoisomerase I in cultured L1210 cells.
Results of filter elution assays of lesions produced in the DNA of cultured L1210 cells by the antineoplastic alkaloid camptothecin support the notion that topoisomerase I is an intracellular target of this drug. One to 10 microM camptothecin induced DNA single-strand, but not double-strand, breaks when incubated with intact cells or with their isolated nuclei. Approximately one half of the strand breakage was protein concealed, as judged by filter elution. Camptothecin-induced, protein-concealed DNA strand breaks disappeared rapidly after drug removal. DNA-protein cross-links were generated by camptothecin with frequencies approximately equal to those of protein-concealed DNA strand breaks. It is likely that camptothecin can inhibit topoisomerase I in intact cells in a manner similar to that in which other antineoplastic agents such as amsacrine or teniposide inhibit topoisomerase II. DNA-breaking lesions other than those resulting from trapped topoisomerase I-DNA complexes may also be generated by camptothecin. The yields of DNA strand breaks induced by camptothecin, amsacrine, or teniposide were approximately doubled when cells were incubated for 16 h with 3-aminobenzamide, an inhibitor of poly(ADP ribosylation) of proteins, prior to 1-h exposure to the antineoplastic compounds. 3-Aminobenzamide also enhanced the cytotoxic action of camptothecin, amsacrine, and teniposide. These results suggest that protein-concealed strand breaks can be lethal lesions and that intracellular topoisomerase I and II activity may be regulated coordinately through poly(ADP ribosylation).


Effects of external application of compound Qingbi granules on acute gouty arthritis with dampness-heat syndrome: a randomized controlled trial.
The use of anti-inflammatory and analgesic drugs such as nonsteroidal anti-inflammatory drugs(NSAIDs) for treating acute gout has limitations, such as adverse reactions in the gastrointestinal tract and toxicity in the liver, kidney, and heart. Hence, a new safe and effective treatment approach needs to be explored to reduce the use of anti-inflammatory and analgesic drugs, incidence of adverse reactions, and patients' burden. This randomized controlled clinical trial aimed to investigate the clinical efficacy and safety of the external application of compound Qingbi granules (CQBG) in treating acute gouty arthritis(AGA), providing evidence for designing a safe, effective, and optimized protocol for AGA comprehensive treatment. A total of 90 patients in line with the diagnostic standard of AGA were recruited and randomly divided into control, T1, and T2 groups (30 in each group). All the participators in the three groups all received Western-medicine-basic treatment (low-purine diet, drinking water more than 2000 mL/days, oral loxoprofen, and NAHCO3). Besides, the T1 group received an external application of diclofenac diethylamine emulgel, while the T2 group received an external application of CQBG. The participants in the control group received single-use Western-medicine-basic treatment. With a treatment course of 7 days and a follow-up of 7 days, the three groups were compared in terms of primary outcome indicators, including swelling, pain improvement, and change in pain duration and secondary outcome indicators, including serum C-reactive protein (CRP) level, uric acid (UA) level, and change in the thickness of the inflammatory synovium of joints under ultrasound. Meanwhile, the safety of the protocol was evaluated. The three groups of patients had no apparent differences in age, body mass index, history of gout, complications, and so on before recruitment. A comparison between pretreatment and post-treatment revealed remarkable reductions in the arthralgia visual analog scale score(VAS) and the swelling score in the three groups after the treatment and the improvements in the T2 group were more significant than those in the T1 and control groups (P < 0.05). Regarding the onset time of pain improvement and pain duration, the T2 group had more significant efficacy compared with the other two groups (P < 0.05). The serum CRP and blood UA levels in the three groups significantly decreased after the treatment, but with no significant intergroup difference. The improvement in the thickness of the inflammatory synovium in joints tested by ultrasound was more significant in the T2 group than in the control group (P < 0.05). For safety evaluations, no significant difference in the incidence of adverse events was found. The external application of CQBG combined with Western-medicine-basic treatment in patients with AGA improved arthralgia and swelling, shortened the period of taking NSAIDs, and reduced the levels of CRP and serum UA. Its therapeutic effect was significantly better than the effect of single-use Western-medicine-basic treatment. The study provided evidence for the clinical application of CQBG combined with Western medicine in treating AGA. ChiCTR, ChiCTR1800018020. Registered 27 August 2018, https://www.chictr.org.cn/showproj.aspx?proj=27138.


Atomic Layer Deposition of Space-Efficient SnO2 Underlayers for BiVO4 Host-Guest Architectures for Photoassisted Water Splitting.
Bismuth vanadate (BiVO4 ) is promising for solar-assisted water splitting. The performance of BiVO4 is limited by charge separation for >70 nm films or by light harvesting for <700 nm films. To resolve this mismatch, host-guest architectures use thin film coatings on 3D scaffolds. Recombination, however, is exacerbated at the extended host-guest interface. Underlayers are used to limit this recombination with a host-underlayer-guest series. Such underlayers consume precious pore volume where typical SnO2 underlayers are optimized with 65-80 nm. In this study, conformal and ultrathin SnO2 underlayers with low defect density are produced by atomic layer deposition (ALD). This shifts the optimized thickness to just 8 nm with significantly improved space efficiency. The materials chemistry thus determines the dimension optimization. Lastly, host-guest architectures are shown to have an applied bias photon-to-charge efficiency of 0.71 %, a new record for a photoanode absorber prepared by ALD.


Association between red cell distribution width and response to methotrexate in rheumatoid arthritis.
Red cell distribution width (RDW) is an unconventional biomarker of inflammation. We aimed to explore its role as a predictor of treatment response in rheumatoid arthritis (RA). Eighty-two RA patients (55 females), median age [interquartile range] 63 years [52-69], were selected by scanning the medical records of a rheumatology clinic, to analyze the associations between baseline RDW, disease activity scores and inflammatory markers, as well as the relationship between RDW changes following methotrexate (MTX) and treatment response. The lower the median baseline RDW, the greater were the chances of a positive EULAR response at three months, 13.5% [13.0-14.4] being among those with good response, vs 14.0% [13.2-14.7] and 14.2% [13.5- 16.0] (p=0.009) among those with moderate and poor response, respectively. MTX treatment was followed by a significant RDW increase (p<0.0001). The increase of RDW was greater among patients with good EULAR response, becoming progressively smaller in cases with moderate and poor response (1.0% [0.4-1.4] vs. 0.7 [0.1-2.0] vs. 0.3 [-0.1-0.8]; p=0.03). RDW is a strong predictor of early response to MTX in RA. RDW significantly increases after MTX initiation in parallel to treatment response, suggesting a role as a marker of MTX effectiveness.


Mechanism of stimulation of ornithine decarboxylase activity in transformed mouse fibroblasts.
Stimulation of confluent serum-starved SV-3T3 cells by serum produces a transient enhancement of ornithine decarboxylase activity which fluctuates at least 20-fold over a 12-h period. The mechanism by which this fluctuation is produced was investigated. Two techniques for assaying the amount of enzyme protein in these cells were utilized: radioimmunoassay and titration with alpha-(difluoromethyl) [5-3H]ornithine. The radioimmunoassay was carried out by using a specific antiserum prepared in rabbits against homogeneous mouse kidney ornithine decarboxylase and by using alpha-(difluoromethyl) [5-3H]ornithine-labeled kidney ornithine decarboxylase as the tracer ligand. An exact correlation between the amount of enzyme protein and the amount of enzyme activity was seen during the rise and fall of ornithine decarboxylase activity after serum stimulation. Similarly, the amount of protein which was labeled covalently by reaction with alpha-(difluoromethyl) [5-3H]ornithine (a specific enzyme-activated irreversible inhibitor) correlated with the enzyme activity. Investigation of the protein labeled in this way by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate indicated that no change in the size of the protein which had a molecular weight of 53 000 occurred during this time period. These results indicate the alteration in ornithine decarboxylase activity can be accounted for entirely by changes in the amount of enzyme protein rather than by posttranslational modifications, activators, or inhibitors.


The vascular biology of atherosclerosis and imaging targets.
The growing worldwide health challenge of atherosclerosis, together with advances in imaging technologies, have stimulated considerable interest in novel approaches to gauging this disease. The last several decades have witnessed a burgeoning in understanding of the molecular pathways involved in atherogenesis, lesion progression, and the mechanisms underlying the complications of human atherosclerotic plaques. The imaging of atherosclerosis is reaching beyond anatomy to encompass assessment of aspects of plaque biology related to the pathogenesis and complication of the disease. The harnessing of these biologic insights promises to provide a plethora of new targets for molecular imaging of atherosclerosis. The goals for the years to come must include translation of the experimental work to visualization of these appealing biologic targets in humans.


The role of cavitation in liposome formation.
Liposome size is a vital parameter of many quantitative biophysical studies. Sonication, or exposure to ultrasound, is used widely to manufacture artificial liposomes, yet little is known about the mechanism by which liposomes are affected by ultrasound. Cavitation, or the oscillation of small gas bubbles in a pressure-varying field, has been shown to be responsible for many biophysical effects of ultrasound on cells. In this study, we correlate the presence and type of cavitation with a decrease in liposome size. Aqueous lipid suspensions surrounding a hydrophone were exposed to various intensities of ultrasound and hydrostatic pressures before measuring their size distribution with dynamic light scattering. As expected, increasing ultrasound intensity at atmospheric pressure decreased the average liposome diameter. The presence of collapse cavitation was manifested in the acoustic spectrum at high ultrasonic intensities. Increasing hydrostatic pressure was shown to inhibit the presence of collapse cavitation. Collapse cavitation, however, did not correlate with decreases in liposome size, as changes in size still occurred when collapse cavitation was inhibited either by lowering ultrasound intensity or by increasing static pressure. We propose a mechanism whereby stable cavitation, another type of cavitation present in sound fields, causes fluid shearing of liposomes and reduction of liposome size. A mathematical model was developed based on the Rayleigh-Plesset equation of bubble dynamics and principles of acoustic microstreaming to estimate the shear field magnitude around an oscillating bubble. This model predicts the ultrasound intensities and pressures needed to create shear fields sufficient to cause liposome size change, and correlates well with our experimental data.


Ubiquinones have surface-active properties suited to transport electrons and protons across membranes.
Surface-active properties of ubiquinones and ubiquinols have been investigated by monomolecular-film techniques. Stable monolayers are formed at an air/water interface by the fully oxidized and reduced forms of the coenzyme; collapse pressures and hence stability of the films tend to increase with decreasing length of the isoprenoid side chain and films of the reduced coenzymes are more stable than those of their oxidized counterparts. Ubiquinone with a side chain of two isoprenoid units does not form stable monolayers at the air/water interface. Mixed monolayers of ubiquinol-10 or ubiquinone-10 with 1,2-dimyristoyl phosphatidylcholine, soya phosphatidylcholine and diphosphatidylglycerol do not exhibit ideal mixing characteristics. At surface pressures less than the collapse pressure of pure ubiquinone-10 monolayers (approx. 12mN.m(-1)) the isoprenoid chain is located substantially within the region occupied by the fatty acyl residues of the phospholipids. With increasing surface pressure the ubiquinones and their fully reduced equivalents are progressively squeezed out from between the phospholipid molecules until, at a pressure of about 35mN.m(-1), the film has surface properties consistent with that of the pure phospholipid monolayer. This suggests that the ubiquinone(ol) forms a separate phase overlying the phospholipid monolayer. The implications of this energetically poised situation, where the quinone(ol) is just able to penetrate the phospholipid film, are considered in terms of the function of ubiquinone(ol) as electron and proton carriers of energy-transducing membranes.


Ralstonia solanacearum Type III Effector RipAY Is a Glutathione-Degrading Enzyme That Is Activated by Plant Cytosolic Thioredoxins and Suppresses Plant Immunity.
The plant pathogen Ralstonia solanacearum uses a large repertoire of type III effector proteins to succeed in infection. To clarify the function of effector proteins in host eukaryote cells, we expressed effectors in yeast cells and identified seven effector proteins that interfere with yeast growth. One of the effector proteins, RipAY, was found to share homology with the ChaC family proteins that function as γ-glutamyl cyclotransferases, which degrade glutathione (GSH), a tripeptide that plays important roles in the plant immune system. RipAY significantly inhibited yeast growth and simultaneously induced rapid GSH depletion when expressed in yeast cells. The in vitro GSH degradation activity of RipAY is specifically activated by eukaryotic factors in the yeast and plant extracts. Biochemical purification of the yeast protein identified that RipAY is activated by thioredoxin TRX2. On the other hand, RipAY was not activated by bacterial thioredoxins. Interestingly, RipAY was activated by plant h-type thioredoxins that exist in large amounts in the plant cytosol, but not by chloroplastic m-, f-, x-, y- and z-type thioredoxins, in a thiol-independent manner. The transient expression of RipAY decreased the GSH level in plant cells and affected the flg22-triggered production of reactive oxygen species (ROS) and expression of pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) marker genes in Nicotiana benthamiana leaves. These results indicate that RipAY is activated by host cytosolic thioredoxins and degrades GSH specifically in plant cells to suppress plant immunity. Ralstonia solanacearum is the causal agent of bacterial wilt disease of plants. This pathogen injects virulence effector proteins into host cells to suppress disease resistance responses of plants. In this article, we report a biochemical activity of R. solanacearum effector protein RipAY. RipAY can degrade GSH, a tripeptide that plays important roles in the plant immune system, with its γ-glutamyl cyclotransferase activity. The high GSH degradation activity of RipAY is considered to be a good weapon for this bacterium to suppress plant immunity. However, GSH also plays important roles in bacterial tolerance to various stresses and growth. Interestingly, RipAY has an excellent safety mechanism to prevent unwanted firing of its enzyme activity in bacterial cells because RipAY is specifically activated by host eukaryotic thioredoxins. This study also reveals a novel host plant protein acting as a molecular switch for effector activation.


Intrinsic and Extrinsic Factors Affecting Skin Surface pH.
The body skin pH can usually vary from 4.0 to 7.0 depending on location. The presence of natural acidic compounds on the skin surface helps maintain the skin's physicochemical properties as well as its protective functions. Since the slightly acidic pH of the skin is extremely important for the skin's protective function, the skin is widely known as "acid mantel." Factors such as age, race, gender, body sites, biochemical differences, and even washing affect the pH of the stratum corneum. Recent clinical studies using an emollient-base finish product using the traditional way of measuring skin pH produced results that indicated an apparent increase in skin pH. The apparent pH increase with these products is most probably an artifact of the skin pH measurement technique. Our findings show that certain petrolatum-based emollients and components could create a protective barrier and help maintain the healthy acidity of the skin. Our work provides new evidence of emollients helping to stabilize skin pH in its natural balanced state rather than affecting it. This new learning should be taken into consideration by other researchers in the area of skin pH as well as in clinical studies to avoid misleading results.


Performance of Next-Generation Sequencing for the Detection of Microsatellite Instability in Colorectal Cancer With Deficient DNA Mismatch Repair.
Next-generation sequencing (NGS) was recently approved by the United States Food and Drug Administration to detect microsatellite instability (MSI) arising from defective mismatch repair (dMMR) in patients with metastatic colorectal cancer (mCRC) before treatment with immune checkpoint inhibitors (ICI). In this study, we aimed to evaluate and improve the performance of NGS to identify MSI in CRC, especially dMMR mCRC treated with ICI. CRC samples used in this post hoc study were reassessed centrally for MSI and dMMR status using the reference methods of pentaplex polymerase chain reaction and immunohistochemistry. Whole-exome sequencing (WES) was used to evaluate MSISensor, the Food and Drug Administration-approved and NGS-based method for assessment of MSI. This was performed in (1) a prospective, multicenter cohort of 102 patients with mCRC (C1; 25 dMMR/MSI, 24 treated with ICI) from clinical trials NCT02840604 and NCT033501260, (2) an independent retrospective, multicenter cohort of 113 patients (C2; 25 mCRC, 88 non-mCRC, all dMMR/MSI untreated with ICI), and (3) a publicly available series of 118 patients with CRC from The Cancer Genome Atlas (C3; 51 dMMR/MSI). A new NGS-based algorithm, namely MSICare, was developed. Its performance for assessment of MSI was compared with MSISensor in C1, C2, and C3 at the exome level or after downsampling sequencing data to the MSK-IMPACT gene panel. MSICare was validated in an additional retrospective, multicenter cohort (C4) of 152 patients with new CRC (137 dMMR/MSI) enriched in tumors deficient in MSH6 (n = 35) and PMS2 (n = 9) after targeted sequencing of samples with an optimized set of microsatellite markers (MSIDIAG). At the exome level, MSISensor was highly specific but failed to diagnose MSI in 16% of MSI/dMMR mCRC from C1 (4 of 25; sensitivity, 84%; 95% confidence interval [CI], 63.9%-95.5%), 32% of mCRC (8 of 25; sensitivity, 68%; 95% CI, 46.5%-85.1%), and 9.1% of non-mCRC from C2 (8 of 88; sensitivity, 90.9%; 95% CI, 82.9%-96%), and 9.8% of CRC from C3 (5 of 51; sensitivity, 90.2%; 95% CI, 78.6%-96.7%). Misdiagnosis included 4 mCRCs treated with ICI, of which 3 showed an overall response rate without progression at this date. At the exome level, reevaluation of the MSI genomic signal using MSICare detected 100% of cases with true MSI status among C1 and C2. Further validation of MSICare was obtained in CRC tumors from C3, with 96.1% concordance for MSI status. Whereas misdiagnosis with MSISensor even increased when analyzing downsampled WES data from C1 and C2 with microsatellite markers restricted to the MSK-IMPACT gene panel (sensitivity, 72.5%; 95% CI, 64.2%-79.7%), particularly in the MSH6-deficient setting, MSICare sensitivity and specificity remained optimal (100%). Similar results were obtained with MSICare after targeted NGS of tumors from C4 with the optimized microsatellite panel MSIDIAG (sensitivity, 99.3%; 95% CI, 96%-100%; specificity, 100%). In contrast to MSISensor, the new MSICare test we propose performs at least as efficiently as the reference method, MSI polymerase chain reaction, to detect MSI in CRC regardless of the defective MMR protein under both WES and targeted NGS conditions. We suggest MSICare may rapidly become a reference method for NGS-based testing of MSI in CRC, especially in mCRC, where accurate MSI status is required before the prescription of ICI.