bluuebunny/crossref_metadata_2025 huggingface dataset:
Somatosensory function in old age
Authors: Heft M. W. , Robinson M. E.
Published: January 2017

Sensory decline is viewed as an inevitable consequence of the ageing process. However, reports of declines have not been a consistent finding across the sensory systems. Reports from psychophysical studies indicate that the most common declines with ageing are in vision and audition and, to a lesser degree, olfaction and gustation. Findings for the somatosensory system (mechanoreception, warming and cooling thermoreception and pain) are less conclusive. Factors that contribute to individual differences in sensory ratings beyond chronological ageing include stimulus factors including stimulus type and body location, response measures and instructions, systemic disease that may affect the peripheral or central nervous system and environmental factors that may affect the skin integrity.
Stripe Rust Resistance in Roegneria kamoji (Poaceae: Triticeae) and its Genetic Analysis
Authors: Zhao Fu‐Qiang , Li Qi , Chen Guo‐Yue , Li Jun , Kang Hou‐Yang , Wang Yi , Fan Xing , Sha Li‐Na , Zhou Yong‐Hong , Zhang Hai‐Qin
Published: January 2017

The Roegneria kamoji accession ZY 1007 was resistant to the mixed predominant races of Puccinia striiformis f.sp. tritici (Pst) in China based on field tests at adult‐plant stage. The seedling resistance evaluation of ZY 1007 showed that it was resistant to stripe rust physiological strains CYR29, CYR33 and PST‐V26, which were the predominant races of Pst in China. The female parent R. kamoji cv. Gansi No.1 (susceptible to Pst) was crossed with ZY 1007 (resistant to Pst). Parents, F1 and F2 populations were tested in a field inoculated with the mixed urediniospores. ZY 1007 and all the observed 11 F1 hybrid plants were resistant, while plants of Gansi No.1 were susceptible. Among the 221 F2 plants, 168 plants were resistant and 53 were susceptible, and the segregation of resistant and susceptible plants fits 3R:1S ratio (χ2 = 0.074, P > 0.75). It confirmed that the resistance of stripe rust in ZY 1007 was controlled by a single dominant gene and temporarily designated as YrK1007.
Comparative DNA sequence analyses of Pyramimonas parkeae (Prasinophyceae) chloroplast genomes
Authors: Satjarak Anchittha , Graham Linda E.
Published: January 2017

Prasinophytes form a paraphyletic assemblage of early diverging green algae, which have the potential to reveal the traits of the last common ancestor of the main two green lineages: (i) chlorophyte algae and (ii) streptophyte algae. Understanding the genetic composition of prasinophyte algae is fundamental to understanding the diversification and evolutionary processes that may have occurred in both green lineages. In this study, we sequenced the chloroplast genome of Pyramimonas parkeae NIES254 and compared it with that of P. parkeae CCMP726, the only other fully sequenced P. parkeae chloroplast genome. The results revealed that P. parkeae chloroplast genomes are surprisingly variable. The chloroplast genome of NIES254 was larger than that of CCMP726 by 3,204 bp, the NIES254 large single copy was 288 bp longer, the small single copy was 5,088 bp longer, and the IR was 1,086 bp shorter than that of CCMP726. Similarity values of the two strains were almost zero in four large hot spot regions. Finally, the strains differed in copy number for three protein‐coding genes: ycf20, psaC, and ndhE. Phylogenetic analyses using 16S and 18S rDNA and rbcL sequences resolved a clade consisting of these two P. parkeae strains and a clade consisting of these plus other Pyramimonas isolates. These results are consistent with past studies indicating that prasinophyte chloroplast genomes display a higher level of variation than is commonly found among land plants. Consequently, prasinophyte chloroplast genomes may be less useful for inferring the early history of Viridiplantae than has been the case for land plant diversification.
Hepatitis B surface antigen loss and clinical outcomes between HBeAg‐negative cirrhosis patients who discontinued or continued nucleoside analogue therapy
Authors: Hung C.‐H. , Wang J.‐H. , Lu S.‐N. , Hu T.‐H. , Lee C.‐M. , Chen C.‐H.
Published: January 2017

We investigated the incidence and predictors of post‐treatment hepatitis B virus (HBV) relapse and hepatitis B surface antigen (HBsAg) loss. After cessation of nucleoside analogue (NA) treatment in hepatitis B e antigen (HBeAg)‐negative patients with cirrhosis. The rates of HBsAg loss and hepatocellular carcinoma (HCC) development in HBeAg‐negative patients with cirrhosis who continued NA treatment were compared with those who discontinued treatment. Patients with compensated cirrhosis who had discontinued NA treatment for at least 12 months (discontinuing group; n=73) and patients who continued entecavir treatment for at least 4 years (continuing group; n=158) were recruited. Serum HBsAg levels were analysed at the end of treatment (discontinuing group) or at 2.5‐3 years of treatment (continuing group). In the discontinuing group, the 6‐year cumulative incidence of post‐treatment virological relapse and HBsAg loss were 56.3% and 46.7%, respectively. The end‐of‐treatment HBsAg level of 300 IU/mL was a cut‐off value for subsequent post‐treatment HBsAg loss and sustained response. In the continuing group, HBsAg loss occurred in five of 158 patients. Cox regression analysis showed that HBsAg levels in the discontinuing group were independent predictors for HBsAg loss in all patients and 104 propensity score (PS)‐matched patients. There was no significant difference in HCC development between the groups in all patients and 104 PS‐matched patients. Two patients experienced post‐treatment alanine aminotransferase flare with hepatic decompensation, and neither of them died after retreatment. In conclusion, HBeAg‐negative patients with cirrhosis who discontinued NA treatment might have a higher rate of HBsAg loss and their risk of developing HCC did not increase compared with those who continued entecavir treatment.
Insufficient levels of the nrdAB‐encoded ribonucleotide reductase underlie the severe growth defect of the Δhda E. coli strain
Authors: Babu Vignesh M. P. , Itsko Mark , Baxter Jamie C. , Schaaper Roel M. , Sutton Mark D.
Published: January 2017

The ATP‐bound form of the Escherichia coli DnaA replication initiator protein remodels the chromosomal origin of replication, oriC, to load the replicative helicase. The primary mechanism for regulating the activity of DnaA involves the Hda and β clamp proteins, which act together to dramatically stimulate the intrinsic DNA‐dependent ATPase activity of DnaA via a process termed Regulatory Inactivation of DnaA. In addition to hyperinitiation, strains lacking hda function also exhibit cold sensitive growth at 30°C. Strains impaired for the other regulators of initiation (i.e., ΔseqA or ΔdatA) fail to exhibit cold sensitivity. The goal of this study was to gain insight into why loss of hda function impedes growth. We used a genetic approach to isolate 9 suppressors of Δhda cold sensitivity, and characterized the mechanistic basis by which these suppressors alleviated Δhda cold sensitivity. Taken together, our results provide strong support for the view that the fundamental defect associated with Δhda is diminished levels of DNA precursors, particularly dGTP and dATP. We discuss possible mechanisms by which the suppressors identified here may regulate dNTP pool size, as well as similarities in phenotypes between the Δhda strain and hda+ strains exposed to the ribonucleotide reductase inhibitor hydroxyurea.
Relationship of serum haemojuvelin and hepcidin levels with iron level and erythropoietin requirement in prevalent hepatitis C virus positive haemodialysis patients
Authors: El Said Heba W. , Abou Seif Khaled H. , Ahmed Yasser S. , Abou Elleil Hesham A. , El Said Tamer W. , Behairy Maha A. , Mohamed Mohamed M. , Ahmed Fatma A.
Published: January 2017

Aim

Iron overload is frequently reported in haemodialysis (HD) patients particularly those with chronic hepatitis C virus (HCV) infection. Soluble haemojuvelin (sHJV) has recently emerged as one of the significant regulators of iron homeostasis and hepcidin expression. The aim of the present study was to evaluate the potential associations of sHJV and hepcidin with inflammation, iron parameters and erythropoietin requirement in prevalent HD patients with HCV.

Methods

Serum sHJV and hepcidin were measured in 60 prevalent HD patients with [group I (n = 30)] and without [group II (n = 30)] HCV, and controls (n = 30) by enzyme‐linked immunosorbent assay. Parameters related to anaemia, iron metabolism, inflammation, sHJV and hepcidin were measured.

Results

Serum hepcidin in HCV positive versus negative groups was 89.40 ± 46.08 ng/mL and 224.1 ± 72.36 ng/mL, P = 0.000, respectively, while sHJV was 245 ± 1.338 ng/mL and 254 ± 0.762 ng/mL, P = 0.147, respectively in positive versus negative patients. In group I, hepcidin correlated with serum ferritin (r = −0.512 P = 0.005) and transferrin saturation (TSAT%) (r = 0.572, P = 0.000) and sHJV correlated with ferritin (r = 0.40, P 0.000), TSAT% (r = 0.450, P = 0.002) and a significant correlation also existed between sHJV and hepcidin (r = −0.259, P = 0.045). In the regression analysis, ferritin and TSAT% were able to predict sHJV; (standardized β = 0.52, P 0.001) and (standardized β = 0.48, P 0.010). Ferritin and sHJV were also able to predict hepcidin (standardized β = 0.627, P = 0.006) and (standardized β = 0.300, P = 0.007) in group I.

Conclusion

Soluble haemojuvelin levels seem to be associated with iron overload parameters and hepcidin levels in HCV positive HD patients.
Photo‐co‐carcinogenesis of Topically Applied Retinyl Palmitate in SKH‐1 Hairless Mice
Authors: Boudreau Mary D. , Beland Frederick A. , Felton Robert P. , Fu Peter P. , Howard Paul C. , Mellick Paul W. , Thorn Brett T. , Olson Greg R.
Published: January 2017

Cosmetic products that contain retinyl palmitate are popular as antiaging skin treatments; however, recent studies suggest a risk for enhanced skin tumor development with topical retinyl palmitate applications and exposure to solar ultraviolet radiation (UVR). In this study, we investigated the potential of retinyl palmitate to enhanceUVR‐induced photo‐co‐carcinogenesis. Groups of 36 male and 36 femaleSKH‐1 hairless mice were exposed to simulated solar light (SSL) and treated with the control cream or creams containing retinyl palmitate, 5 days per week for 40 weeks. Other groups of mice were exposed toSSLand received no cream treatment or received cream treatments and were exposed to ultraviolet‐A or ultraviolet‐B. Mice were monitored for the development of skin tumors, and the incidences and multiplicities of squamous cell neoplasia were determined by histopathology. In both the absence and presence ofSSL, mice administered the control cream developed skin tumors earlier and had higher incidences and multiplicities of skin squamous cell neoplasms than mice that received no cream treatment. Compared to the control cream groups, mice exposed toSSLand administered the retinyl palmitate creams demonstrated earlier onsets of skin tumors and had increased incidences and multiplicities of squamous cell skin neoplasms.
Influence of demographic and clinical characteristics of actinic keratosis patients on illness perceptions and readiness to increase sun protection behaviours: an exploratory study
Authors: Akarsu Sevgi , Ozbagcivan Ozlem , Ilknur Turna , Semiz Fatma , Fetil Emel
Published: January 2017

Background/purpose

There are no data comprehensively investigating the key beliefs and sun protection behaviours in patients with actinic keratosis (AK). Our aim was to determine and correlate the AK perceptions and readiness to increase sun protection (RISP) behaviours.

Methods

AK patients’ (n = 200) demographics and clinical findings were recorded. Participants completed questionnaires (Illness Perception Questionnaire‐Revised and RISP behaviours scales).

Results

The mean scores of giving up sunbathing of the patients aged <65 years, and giving up sunbathing and staying in the shade behaviours of females were higher. As the level of education increased, while the mean scores of giving up sunbathing decreased, using sunscreen scores increased. The mean scores of giving up sunbathing, using sunscreen and wearing protective clothing of those with the history of non‐skin cancer (had cancer other than of the skin) were lower. The readiness to increase using sunscreen was found to be higher in patients with more chronic timeline beliefs, more positive perceived personal control and blaming sunlight more, and the wearing protective clothing was found to be higher in those with more cyclical timeline beliefs.

Conclusion

AK patients, especially the subjects aged ≥65 years, have non‐skin cancer history, and male gender should be informed to apply sun‐protective behaviours.
The effect of Echinococcus granulosus on spleen cells and TGF‐β expression in the peripheral blood of BALB/c mice
Authors: Yin S. , Chen X. , Zhang J. , Xu F. , Fang H. , Hou J. , Zhang X. , Wu X. , Chen X.
Published: January 2017

Cystic echinococcosis (CE) caused by the cestode Echinococcus granulosus (E. granulosus) is a zoonotic parasitic disease. The effective immune evasion mechanisms of E. granulosus allow it to parasitize its hosts. However, the status of the innate and adaptive immune cells and their contributions to E. granulosus progression remain poorly understood. In this study, we aimed to determine the impact of E. granulosus infection on T cells, NK cell responses and TGF‐β expression during the early infection phase in BALB/c mice. In E. granulosus infections, there was an increasing tendency in the percentage of CD4+CD25+ T cells and CD4+Foxp3+ T cells and peripheral blood TGF‐β levels and relative expression of the Foxp3 gene. Moreover, there were a decreasing tendency in the percentage of NK cells and NK cell cytotoxicity and the expression of NKG2D on NK cells. The TGF‐β1/Smad pathway was activated by E. granulosus in mice. Above results can be reversed by the inhibitor SB‐525334 (potent activin receptor‐like kinase 5 inhibitor). These results suggest that the TGF‐β/Smad pathway plays an important role in changes of T‐cell or NK cell responses. These results may contribute to revealing the preliminary molecular mechanisms in establishing hydatid infection.
Confirmed mixed bird–insect pollination system of Scrophularia trifoliata L., a Tyrrhenian species with corolla spots
Authors: Navarro‐Pérez M. L. , López J. , Rodríguez‐Riaño T. , Bacchetta G. , de Miguel Gordillo C. , Ortega‐Olivencia A.
Published: January 2017

Both bird and mixed vertebrate–insect (MVI) pollination systems are very rare in Europe and the Mediterranean region. Because MVI can ensure reproduction over a wider range of environmental conditions than when insects are the sole pollinators, under certain circumstances such systems are highly advantageous to plants. Here, we investigated the pollination and some reproductive traits of the Tyrrhenian Scrophularia trifoliata, the only species of the genus possessing two showy dark spots inside the corolla, for which MVI pollination system had been inferred on the basis of limited censuses.

We conducted field experiments to study MVI pollination and some reproductive traits and elucidate the role of corolla spots, analysing their ultraviolet pattern, histology and pigments versus the rest of the corolla.

The primary pollinators were wasps and passerine birds. Corolla spots absorb UV light, present abundant anthocyanins and are histologically identical to the rest of the corolla. Control flowers had higher visitation frequency than flowers without spots. S. trifoliata is self‐compatible, with efficient intrafloral protogyny and herkogamy that prevent self‐pollination but not geitonogamy.

We confirmed the existence of a mixed bird–insect pollination system in S. trifoliata. This system is found in three other Scrophularia species with large, showy flowers – two Mediterranean (S. grandiflora and S. sambucifolia) and one Macaronesian (S. calliantha). Unlike those species, S. trifoliata has two large spots inside the corolla. These likely operate as nectar guides and their dark colouration is related to abundant anthocyanin content.
Falls, risk factors and fear of falling among persons older than 65 years of age
Authors: Gazibara Tatjana , Kurtagic Ilma , Kisic‐Tepavcevic Darija , Nurkovic Selmina , Kovacevic Nikolina , Gazibara Teodora , Pekmezovic Tatjana
Published: January 2017

Aim

Falling represents a major public health problem among older persons because it leads to premature mortality, loss of independence, and placement in assisted‐living facilities. The purpose of this study was to assess the main features and risks for falls among persons older than 65 years of age as well as to quantify their fear of falling.

Methods

A total of 354 persons older than 65 years of age were recruited at a community health centre. Characteristics of the most recent fall were obtained through detailed interviews with study participants. The Falls Efficacy Scale was used to quantify fear of falling.

Results

Frequency of falling was 15.8%. Falls occurred most often while walking (49%). One‐half of fallers (49.1%) sustained an injury. Head haematomas and soft tissues contusions were the most common consequences of falls. The average Falls Efficacy Scale score was significantly higher in fallers (
P
= 0.001). Multiple logistic regression analysis showed that having a fear of falling (odds ratio = 4.14, 95% confidence interval: 1.22–14.08,
P
= 0.02) and being a woman (odds ratio = 2.10, 95% confidence interval: 0.97–4.53,
P
= 0.05) were independent risk factors for falling among older persons.

Conclusion

The frequency of falls among older people was similar to those in other populations. These results could be used to help select older persons who should be enrolled in fall prevention programmes.
The role of social relationships among elderly community‐dwelling and nursing‐home residents: findings from a quality of life study
Authors: Scocco Paolo , Nassuato Mario
Published: January 2017

Aim

In Western countries, older adults' needs are often managed through institutionalization. Based on the assumption that quality of life, particularly social relationships, may be perceived differently according to residential setting, the aims of this study were to compare World Health Organization Quality of Life brief version (WHOQOL‐BREF) scores of elderly community‐dwelling residents and nursing home residents.

Methods

A sample of 207 older adults (135 community‐dwelling residents, 72 nursing home residents) was evaluated with Mini‐Mental State Examination (MMSE), WHOQOL‐BREF, and Geriatric Depression Scale (GDS).

Results

Nursing home residents achieved lower WHOQOL‐BREF scores on the physical health scale only (P = 0.002). In a linear regression model, physical score correlated negatively with GDS score (P = 0.0001) and Mini‐Mental State Examination score (P = 0.04), but positively with male gender (P = 0.02) and community‐dwelling residence (P = 0.001); psychological score correlated negatively with GDS score (P = 0.0001) and being married (P = 0.03), but positively with male gender (P = 0.009) and being unmarried (P = 0.03). The social relationships score correlated negatively with the GDS score (P = 0.0001) and male gender (P = 0.02), but positively with high education level (P = 0.04). The environment score negatively correlated with GDS score (P = 0.0001). In a logistic regression model, living in a nursing home correlated with female gender (P = 0.001), age (P = 0.0001), a lower physical score (P = 0.0001), and a higher social relationships score (P = 0.02).

Conclusion

Depressive symptoms correlated with low scores in all WHOQOL‐BREF domains. The variables that correlated with living conditions in a nursing home were older age, male gender, lower physical domain scores, and higher social relationship scores. Opportunities for socialization in nursing homes may thus improve perception of quality of life in this domain.
Do spouse caregivers of young and older persons with dementia have different needs? A comparative study
Authors: Wawrziczny Emilie , Pasquier Florence , Ducharme Francine , Kergoat Marie‐Jeanne , Antoine Pascal
Published: January 2017

Aim

The aim of this study was to explore the needs of spouse caregivers of persons with dementia (PWD) and then to compare them based on the PWD's age at disease onset. This data could be used to adapt support programmes to address differences between the two groups.

Method

Thirty‐eight spouse caregivers of persons with late‐onset dementia and 40 spouse caregivers of persons with early‐onset dementia (PEOD) agreed to participate in the study. The mean ± SD age of the PEOD was 57.6 ± 4.0 years, whereas it was 80.9 ± 5.3 years for the persons with late‐onset dementia. Interviews were conducted in the spouse caregivers' homes with only the spouse caregiver. The semi‐structured interviews were based on the French version of the Carers Outcome Agreement Tool. The interviews were analyzed in two steps. The first step was qualitative to identify needs. The data were thematically analyzed using QSR NVivo 10. The second step was quantitative to compare the needs depending on the PWD's age at onset. The comparison between the two groups was performed using the χ2 test.

Results

The results demonstrated that the majority of needs are the same for the two groups of spouse caregivers. All caregivers need to unwind, to stimulate and pay attention to the PWD, to break the isolation, and to be more prepared and confident. However, some differences emerge, with the spouse caregivers of PEOD expressing a greater number of needs. The caregivers of PEOD seem to have a greater need to interact and maintain contacts with other people (P = 0.001), have more general care‐related needs (P = 0.005), require more appropriate care structures (P = 0.037), and need greater assistance with administrative procedures (P = 0.004).

Conclusion

To improve spouse caregivers' well‐being and sense of efficiency, it would be interesting to develop a support programme with a common framework and specific modules depending on the PWD's age at disease onset.
Possible effectiveness of low‐dose pregabalin in a patient suffering from Lewy body disease and chronic pruritus
Authors: Ukai Katsuyuki
Published: January 2017

Many patients with Lewy body disease (LBD) complain of pain, although the detailed mechanism responsible for LBD‐related pain remains to be clarified. Moreover, there does not appear to be any reports about LBD patients suffering from an itching sensation. In this report, the case of an LBD patient who experienced severe itching that persisted for over 6 months, which was treated successfully with low‐dose pregabalin therapy, is described. This is the first study to report the effects of low‐dose pregabalin, which has been used to treat LBD‐related pain, to treat itching in a patient with LBD.
Feelings of uselessness and 3‐year mortality in an Italian community older people: the role of the functional status
Authors: Curzio Olivia , Bernacca Emilia , Bianchi Bruno , Rossi Giuseppe
Published: January 2017

Background

Sense of self‐worth influences the health status of the elderly and may be associated with mortality. The aim of the study was to investigate whether the association between subjective feelings of uselessness and mortality was confounded or modified by functional limitation in non‐institutionalized older people.

Methods

Participants were community‐dwelling older people, aged 70 years and older, who lived in neighbourhoods of Massa and Carrara municipalities in northern Tuscany, Italy. At baseline, 2335 non‐institutionalized older people were assessed with a short self‐administered questionnaire; the analysis included 2132 older persons for whom vital statistical data were available after a 3‐year follow‐up.

Results

The feeling of uselessness was associated with an increased mortality at the 3‐year follow‐up, but only in older people who reported disability problems (adjusted hazard ratio = 1.97, 95% confidence interval = 1.48–2.63, P < 0.0001).

Conclusions

People who reported disability problems and a feeling of uselessness may be vulnerable to an increased risk for poor health outcomes in later life. This study outlined the importance of enquiring about feelings of uselessness, which is a relational variable that is linked to both psychological and physical health status, especially in older people who need help in daily activities.
Klotho attenuated antibody‐mediated porcine endothelial cell activation and injury
Authors: Liu Lu , Gao Hanchao , Hong Chungu , He Chen , Pan Dengke , Dai Yifan , Hara Hidetaka , Cooper David K. C. , Li Zesong , Cai Zhiming , Mou Lisha
Published: January 2017

Long‐term success in pig‐to‐primate xenotransplantation is currently hampered by acute vascular rejection (AVR), characterized by endothelial cell (EC) activation and injury. Klotho has anti‐apoptotic, anti‐inflammatory effects on EC and protects EC against reactive oxygen species, rendering klotho a promising molecule to control AVR. In this study, porcine ECs were pre‐incubated with klotho and then exposed to xenoreactive antibodies and complement. Real‐time PCR revealed that klotho suppressed antibody‐induced pro‐inflammatory gene expression of VCAM‐1 and IL‐1α. NF‐κB activation, IκBα phosphorylation, was also attenuated by klotho administration. Furthermore, klotho induced in porcine EC resistance against complement‐dependent cytotoxicity. Accompanying this change, the binding of IgG and IgM xenoreactive antibodies to porcine EC was decreased and the expression of anti‐inflammatory gene HO‐1 was upregulated. These findings indicated that klotho protein protected porcine EC from activation and injury caused by binding of xenoreactive antibodies and was a promising candidate molecule in a multitransgenic pig strategy for xenotransplantation.
Transplantation of hepatocytes from genetically engineered pigs into baboons
Authors: Iwase Hayato , Liu Hong , Schmelzer Eva , Ezzelarab Mohamed , Wijkstrom Martin , Hara Hidetaka , Lee Whayoung , Singh Jagjit , Long Cassandra , Lagasse Eric , Gerlach Jörg C. , Cooper David K. C. , Gridelli Bruno
Published: January 2017

Background

Some patients with acute or acute‐on‐chronic hepatic failure die before a suitable human liver allograft becomes available. Encouraging results have been achieved in such patients by the transplantation of human hepatocyte progenitor cells from fetal liver tissue. The aim of the study was to explore survival of hepatocytes from genetically engineered pigs after direct injection into the spleen and other selected sites in immunosuppressed baboons to monitor the immune response and the metabolic function and survival of the transplanted hepatocytes.

Methods

Baboons (n=3) were recipients of GTKO/hCD46 pig hepatocytes. All three baboons received anti‐thymocyte globulin (ATG) induction and tapering methylprednisolone. Baboon 1 received maintenance immunosuppressive therapy with tacrolimus and rapamycin. Baboons 2 and 3 received an anti‐CD40mAb/rapamycin‐based regimen that prevents sensitization to pig solid organ grafts. The baboons were euthanized 4 or 5 weeks after hepatocyte transplantation. The baboon immune response was monitored by the measurement of anti‐non‐Gal IgM and IgG antibodies (by flow cytometry) and CFSE‐mixed lymphocyte reaction. Monitoring for hepatocyte survival and function was by (i) real‐time PCR detection of porcine DNA, (ii) real‐time PCR for porcine gene expression, and (iii) pig serum albumin levels (by ELISA). The sites of hepatocyte injection were examined microscopically.

Results

Detection of porcine DNA and porcine gene expression was minimal at all sites of hepatocyte injection. Serum levels of porcine albumen were very low—500‐1000‐fold lower than in baboons with orthotopic pig liver grafts, and approximately 5000‐fold lower than in healthy pigs. No hepatocytes or infiltrating immune cells were seen at any of the injection sites. Two baboons (Baboons 1 and 3) demonstrated a significant increase in anti‐pig IgM and an even greater increase in IgG, indicating sensitization to pig antigens.

Discussion and Conclusions

As a result of this disappointing experience, the following points need to be considered. (i) Were the isolated pig hepatocytes functionally viable? (ii) Are pig hepatocytes more immunogenic than pig hearts, kidneys, artery patch grafts, or islets? (iii) Does injection of pig cells (antigens) into the spleen and/or lymph nodes stimulate a greater immune response than when pig tissues are grafted at other sites? (iv) Did the presence of the recipient's intact liver prevent survival and proliferation of pig hepatocytes? (v) Is pig CD47‐primate SIRP‐α compatibility essential? In conclusion, the transplantation of genetically engineered pig hepatocytes into multiple sites in immunosuppressed baboons was associated with very early graft failure. Considerable further study is required before clinical trials should be undertaken.
Postprandial adjustments in renal phosphate excretion do not involve a gut‐derived phosphaturic factor
Authors: Lee Grace J. , Mossa‐Al Hashimi Lina , Debnam Edward S. , Unwin Robert J. , Marks Joanne
Published: January 2017

New Findings

What is the central question of this study?

Does a previously hypothesized signalling mechanism, believed to detect postprandial increases in intestinal phosphate and that can stimulate the kidneys to rapidly excrete phosphate, operate under physiological conditions?

What is the main finding and its importance?

Contrary to earlier reports, rapid signalling between the small intestine and kidney mediated by a gut‐derived phosphaturic factor in response to a physiological intestinal phosphate load is not supported by the present findings; moreover, hyperphosphataemia and increased parathyroid hormone concentrations are likely to be the underlying factors responsible for the phosphaturia following a supraphysiological intestinal phosphate load.

To date, the role of the small intestine in the regulation of postprandial phosphate homeostasis has remained unclear and controversial. Previous studies have proposed the presence of a gut‐derived phosphaturic factor that acts independently of changes in plasma phosphate concentration or parathyroid hormone (PTH) concentration; however, these early studies used duodenal luminal phosphate concentrations in the molar range, and therefore, the physiological relevance of this is uncertain. In the present study, we used both in vivo and in vitro approaches to investigate the presence of this putative ‘intestinal phosphatonin’. Instillation of 1.3 m phosphate into the duodenum rapidly induced phosphaturia, but in contrast to previous reports, this was associated with significant hyperphosphataemia and elevated PTH concentration; however, there was not the expected decrease in abundance of the renal sodium–phosphate cotransporter NaPi‐IIa. Instillation of a physiological (10 mm) phosphate load had no effect on plasma phosphate concentration, PTH concentration or phosphate excretion. Moreover, phosphate uptake by opossum kidney cells was unaffected after incubation with serosal fluid collected from intestinal segments perfused with different concentrations of phosphate. Taken together, these findings do not support the concept of a gut‐derived phosphaturic factor that can mediate rapid signalling between the gut and kidney, leading to increased urinary phosphate excretion, as part of normal phosphate homeostasis.
Calmodulin and ATP support activity of the Cav1.2 channel through dynamic interactions with the channel
Authors: Minobe Etsuko , Mori Masayuki X. , Kameyama Masaki
Published: January 2017

Key points

Cav1.2 channels maintain activity through interactions with calmodulin (CaM). In this study, activities of the Cav1.2 channel (α1C) and of mutant‐derivatives, C‐terminal deleted (α1CΔ) and α1CΔ linked with CaM (α1CΔCaM), were compared in the inside‐out mode.

α1CΔ with CaM, but not without CaM, and α1CΔCaM were active, suggesting that CaM induced channel activity through a dynamic interaction with the channel, even without the distal C‐tail.

ATP induced α1C activity with CaM and enhanced activity of the mutant channels. Okadaic acid mimicked the effect of ATP on the wildtype but not mutant channels.

These results supported the hypothesis that CaM and ATP maintain activity of Cav1.2 channels through their dynamic interactions. ATP effects involve mechanisms both related and unrelated to channel phosphorylation.

CaM‐linked channels are useful tools for investigating Cav1.2 channels in the inside‐out mode; the fast run‐down is prevented by only ATP and the slow run‐down is nearly absent.

Calmodulin (CaM) plays a critical role in regulation of Cav1.2 Ca2+ channels. CaM binds to the channel directly, maintaining channel activity and regulating it in a Ca2+‐dependent manner. To explore the molecular mechanisms involved, we compared the activity of the wildtype channel (α1C) and mutant derivatives, C‐terminal deleted (α1C∆) and α1C∆ linked to CaM (α1C∆CaM). These were co‐expressed with β2a and α2δ subunits in HEK293 cells. In the inside‐out mode, α1C and α1C∆ showed minimal open‐probabilities in a basic internal solution (run‐down), whereas α1C∆ with CaM and α1C∆CaM maintained detectable channel activity, confirming that CaM was necessary, but not sufficient, for channel activity. Previously, we reported that ATP was required to maintain channel activity of α1C. Unlike α1C, the mutant channels did not require ATP for activation in the early phase (3–5 min). However, α1C∆ with CaM + ATP and α1C∆CaM with ATP maintained activity, even in the late phase (after 7–9 min). These results suggested that CaM and ATP interacted dynamically with the proximal C‐terminal tail of the channel and, thereby, produced channel activity. In addition, okadaic acid, a protein phosphatase inhibitor, could substitute for the effects of ATP on α1C but not on the mutant channels. These results supported the hypothesis that CaM and ATP maintain activity of Cav1.2 channels, further indicating that ATP has dual effects. One maintains phosphorylation of the channel and the other becomes apparent when the distal carboxyl‐terminal tail is removed.
Splashing transients of 2D plasmons launched by swift electrons
Authors: Lin Xiao , Kaminer Ido , Shi Xihang , Gao Fei , Yang Zhaoju , Gao Zhen , Buljan Hrvoje , Joannopoulos John D. , Soljačić Marin , Chen Hongsheng , Zhang Baile
Published: January 2017

Revealing how 2D plasmons emerge and evolve in electron energy–loss spectroscopy (EELS).
Potent and broad HIV-neutralizing antibodies in memory B cells and plasma
Authors: Williams LaTonya D. , Ofek Gilad , Schätzle Sebastian , McDaniel Jonathan R. , Lu Xiaozhi , Nicely Nathan I. , Wu Liming , Lougheed Caleb S. , Bradley Todd , Louder Mark K. , McKee Krisha , Bailer Robert T. , O’Dell Sijy , Georgiev Ivelin S. , Seaman Michael S. , Parks Robert J. , Marshall Dawn J. , Anasti Kara , Yang Guang , Nie Xiaoyan , Tumba Nancy L. , Wiehe Kevin , Wagh Kshitij , Korber Bette , Kepler Thomas B. , Munir Alam S. , Morris Lynn , Kamanga Gift , Cohen Myron S. , Bonsignori Mattia , Xia Shi-Mao , Montefiori David C. , Kelsoe Garnett , Gao Feng , Mascola John R. , Moody M. Anthony , Saunders Kevin O. , Liao Hua-Xin , Tomaras Georgia D. , Georgiou George , Haynes Barton F.
Published: January 2017

Plasma is a source of broadly neutralizing antibodies for recombinant antibodies with enhanced potency and breadth.
Differential Attachment of Salmonella enterica and Enterohemorrhagic Escherichia coli to Alfalfa, Fenugreek, Lettuce, and Tomato Seeds
Authors: Cui Yue , Walcott Ronald , Chen Jinru
Published: January 2017

Vegetable seeds have the potential to disseminate and transmit foodborne bacterial pathogens. This study was undertaken to assess the abilities of selected
Salmonella
and enterohemorrhagic
Escherichia coli
(EHEC) strains to attach to fungicide-treated versus untreated, and intact versus mechanically damaged, seeds of alfalfa, fenugreek, lettuce, and tomato. Surface-sanitized seeds (2 g) were exposed to four individual strains of
Salmonella
or EHEC at 20°C for 5 h. Contaminated seeds were rinsed twice, each with 10 ml of sterilized water, before being soaked overnight in 5 ml of phosphate-buffered saline at 4°C. The seeds were then vortexed vigorously for 1 min, and pathogen populations in seed rinse water and soaking buffer were determined using a standard plate count assay. In general, the
Salmonella
cells had higher attachment ratios than the EHEC cells. Lettuce seeds by unit weight had the highest numbers of attached
Salmonella
or EHEC cells, followed by tomato, alfalfa, and fenugreek seeds. In contrast, individual fenugreek seeds had more attached pathogen cells, followed by lettuce, alfalfa, and tomato seeds. Significantly more
Salmonella
and EHEC cells attached to mechanically damaged seeds than to intact seeds (
P
< 0.05). Although, on average, significantly more
Salmonella
and EHEC cells were recovered from untreated than fungicide-treated seeds (
P
< 0.05), fungicide treatment did not significantly affect the attachment of individual bacterial strains to vegetable seeds (
P
> 0.05), with a few exceptions. This study fills gaps in the current body of literature and helps explain bacterial interactions with vegetable seeds with differing surface characteristics.

IMPORTANCE
Vegetable seeds, specifically sprout seeds, have the potential to disseminate and transmit foodborne bacterial pathogens. This study investigated the interaction between two important bacterial pathogens, i.e.,
Salmonella
and EHEC, and vegetable seeds with differing surface characteristics. This research helps understand whether seed surface structure, integrity, and fungicide treatment affect the interaction between bacterial cells and vegetable seeds.
Novel Aldo-Keto Reductases for the Biocatalytic Conversion of 3-Hydroxybutanal to 1,3-Butanediol: Structural and Biochemical Studies
Authors: Kim Taeho , Flick Robert , Brunzelle Joseph , Singer Alex , Evdokimova Elena , Brown Greg , Joo Jeong Chan , Minasov George A. , Anderson Wayne F. , Mahadevan Radhakrishnan , Savchenko Alexei , Yakunin Alexander F.
Published: January 2017

The nonnatural alcohol 1,3-butanediol (1,3-BDO) is a valuable building block for the synthesis of various polymers. One of the potential pathways for the biosynthesis of 1,3-BDO includes the biotransformation of acetaldehyde to 1,3-BDO via 3-hydroxybutanal (3-HB) using aldolases and aldo-keto reductases (AKRs). This pathway requires an AKR selective for 3-HB, but inactive toward acetaldehyde, so it can be used for one-pot synthesis. In this work, we screened more than 20 purified uncharacterized AKRs for 3-HB reduction and identified 10 enzymes with significant activity and nine proteins with detectable activity. PA1127 fromPseudomonas aeruginosashowed the highest activity and was selected for comparative studies with STM2406 fromSalmonella entericaserovar Typhimurium, for which we have determined the crystal structure. Both AKRs used NADPH as a cofactor, reduced a broad range of aldehydes, and showed low activities toward acetaldehyde. The crystal structures of STM2406 in complex with cacodylate or NADPH revealed the active site with bound molecules of a substrate mimic or cofactor. Site-directed mutagenesis of STM2406 and PA1127 identified the key residues important for the activity against 3-HB and aromatic aldehydes, which include the residues of the substrate-binding pocket and C-terminal loop. Our results revealed that the replacement of the STM2406 Asn65 by Met enhanced the activity and the affinity of this protein toward 3-HB, resulting in a 7-fold increase inkcat/Km. Our work provides further insights into the molecular mechanisms of the substrate selectivity of AKRs and for the rational design of these enzymes toward new substrates.

IMPORTANCEIn this study, we identified several aldo-keto reductases with significant activity in reducing 3-hydroxybutanal to 1,3-butanediol (1,3-BDO), an important commodity chemical. Biochemical and structural studies of these enzymes revealed the key catalytic and substrate-binding residues, including the two structural determinants necessary for high activity in the biosynthesis of 1,3-BDO. This work expands our understanding of the molecular mechanisms of the substrate selectivity of aldo-keto reductases and demonstrates the potential for protein engineering of these enzymes for applications in the biocatalytic production of 1,3-BDO and other valuable chemicals.
Dicer and Argonaute Genes Involved in RNA Interference in the Entomopathogenic Fungus Metarhizium robertsii
Authors: Meng Huimin , Wang Zhangxun , Wang Yulong , Zhu Hong , Huang Bo
Published: January 2017

RNA interference (RNAi) is a gene-silencing mechanism that plays an important role in gene regulation in a number of eukaryotic organisms. Two core components, Dicer and Argonaute, are central in the RNAi machinery. However, the physiological roles of Dicer and Argonaute in the entomopathogenic fungus
Metarhizium robertsii
have remained unclear. Here, the roles of genes encoding Dicer (
M. robertsii
dcl1
[
Mrdcl1
] and
Mrdcl2
) and Argonaute (
Mrago1
and
Mrago2
) proteins in
M. robertsii
were investigated. The results showed that the Dicer-like protein MrDCL2 and Argonaute protein MrAGO1 are the major components of the RNAi process occurring in
M. robertsii
. The Dicer and Argonaute genes were not involved in the regulation of growth and diverse abiotic stress response in
M. robertsii
under the tested conditions. Moreover, our results showed that the Dicer and Argonaute gene mutants demonstrated reduced abilities to produce conidia, compared to the wild type (WT) and the gene-rescued mutant. In particular, the conidial yields in the Δ
dcl2
and Δ
ago1
mutants were reduced by 55.8% and 59.3%, respectively, compared with those from the control strains. Subsequently, for the WT and Δ
dcl2
mutant strains, digital gene expression (DGE) profiling analysis of the stage of mycelium growth and conidiogenesis revealed that modest changes occur in development or metabolism processes, which may explain the reduction in conidiation in the Δ
dcl2
mutant. In addition, we further applied high-throughput sequencing technology to identify small RNAs (sRNAs) that are differentially expressed in the WT and the Δ
dcl2
mutant and found that 4 known microRNA-like small RNAs (milRNAs) and 8 novel milRNAs were
Mrdcl2
dependent in
M. robertsii
.

IMPORTANCE
The identification and characterization of components in RNAi have contributed significantly to our understanding of the mechanism and functions of RNAi in eukaryotes. Here, we found that Dicer and Argonaute genes play an important role in regulating conidiation in
M. robertsii
. Our study also demonstrates that diverse small RNA pathways exist in
M. robertsii
. The study provides a theoretical platform for exploration of the functions of Dicer and Argonaute genes involved in RNAi in fungi.
Making an IMPAKT; Improving care of Chronic Kidney Disease patients in the community through collaborative working and utilizing Information Technology
Authors: Xu Gang , Major Rupert , Shepherd David , Brunskill Nigel
Published: January 2017

Chronic kidney disease (CKD) is a serious long-term condition, which if left untreated causes significant cardiovascular sequele. It is well recognized management of modifiable risk factors, such as blood pressure (BP), can lead to improved long-term outcomes. A novel information technology (IT) solution presents a possible solution to help clinicians in the community identify and manage at risk patients more efficiently.

The IMproving Patient care and Awareness of Kidney disease progression Together (IMPAKT) IT tool was used to identify patients with CKD and uncontrolled hypertension in the community. A CKD nurse utilized the tool at primary care practices to identify patients who warranted potential intervention and disseminated this information to clinical staff. Blood pressure management targets and incidence of coded CKD were used to evaluate the project.

Altogether 48 practices participated in an 18 month project from April 2014, and data from 20 practices, with a total adult population of 121,362, was available for analysis. Two full consecutive QI (Quality Improvement) audit cycles were completed. There was an increase in the mean recorded prevalence of coded CKD patients over the course of the project. Similarly, there was an increase in the percentage of patients with BP been recorded and importantly there was an accompanying significant increase in CKD patients achieving BP targets.

At the end of the project an additional 345 individuals with CKD achieved better blood pressure control. This could potentially prevent 9 cardiovascular events in the CKD group, translating to a cost saving of £320,000 for the 20 practices involved. The most significant change in clinical markers occurred during cycle 1 of the audit, the improvement was maintained throughout cycle 2 of the audit. Our results show the real-life clinical impact of a relatively simple and easy to implement QI project, to help improve outcomes in patients with CKD. This was achieved through more efficient working by targeting of high-risk groups, and improved communication between primary/secondary care. The project could be adapted for other chronic disease conditions. Despite the recorded improvements in blood pressure management, a large proportion of high-risk patients remained above ideal blood pressure, additional interventions in this area need to be explored.

Through collaborative and multi-professional working and utilizing IT resources, we have shown it is possible to deliver measurable and sustainable improvements in blood pressure control for patients with CKD in a real life clinical setting.
Improving physician's hand over among oncology staff using standardized communication tool
Authors: Alolayan Ashwaq , Alkaiyat Mohammad , Ali Yosra , Alshami Mona , Al-Surimi Khaled , Jazieh Abdul-Rahman
Published: January 2017

Cancer patients are frequently admitted to hospital for many reasons. During their hospitalization they are handled by different physicians and other care providers. Maintaining good communication among physicians is essential to assure patient safety and the delivery of quality patient care. Several incidents of miscommunication issues have been reported due to lack of a standardized communication tool for patients' hand over among physicians at our oncology department. Hence, this improvement project aims at assessing the impact of using a standardized communication tool on improving patients' hand over and quality of patient care. A quality improvement team has been formed to address the issue of cancer patients' hand over. We adopted specific hand over tool to be used by physicians. This tool was developed based on well-known and validated communication tool called ISBAR - Identify, Situation, Background, Assessment and Recommendation, which contains pertinent information about the patient's condition. The form should be shared at a specific point in time during the handover process. We monitored the compliance of physician's with this tool over 16 weeks embedded by four ‘purposive’ and ‘sequential’ Plan-Do-Study-Act (PDSA) cycles; where each PDSA cycle was developed based on the challenges faced and lessons learned in each step and the result of the previous PDSA cycle. Physicians compliance rate of using the tool had improved significantly from 45% (baseline) to 100% after the fourth PDSA cycle. Other process measure was measuring acknowledgment of hand over receipt email at two checkpoints at 8:00 – 9:00 a.m. and 4:00 – 5:00 p.m. The project showed that using a standardized handover form as a daily communication method between physicians is a useful idea and feasible to improve cancer patients handover with positive impact on many aspects of healthcare process and outcomes.