Dark oxidation of unsaturated lipids by the photo-oxidized 8-methoxypsoralen.
8-Methoxypsoralen (8-MOP) in ethanol, acetone, benzene, or CC14 is photo-oxidized under UV-irradiation (320-400 nm). Photo-oxidized 8-MOP (O2-8-MOP) is stable in the organic solvents, but it is destructed in water or in liposome suspension. The destruction rate constants are 0.04 s(-1) in water and 0.004 s(-1) in liposome membranes as estimated by the kinetics of the chemiluminescence accompanying the destruction. In course of O2-8-MOP destruction the residues of phospholipid unsaturated fatty acids are oxidized . Generation of the singlet oxygen (1 delta g) by excited 8-MOP is observed neither nor in ethanol. Quantum yield of 1 delta g) formation in CCl4 is less than 3%. A pattern is proposed for 8-MOP-sensitized oxidation of unsaturated lipids proceeding without direct attack of lipids by singlet oxygen.


Ornithogenic vegetation: How significant has the seabird influence been on the Aleutian Island vegetation during the Holocene?
In the Aleutian Islands during the Holocene, terrestrial predators were actually absent; as a result, large seabird colonies thrived along the coasts or across entire islands. Bird guano enriches the soil with nitrogen, which can lead to the formation of highly modified ornithogenic (bird-formed) ecosystems. For a more detailed investigation of avian influence, we reconstructed more than 10,000-year-old vegetation dynamics of the coast of Shemya Island (Near Islands) by pollen analysis. At the initial stages of vegetation development (10,000-4,600 cal year BP), sedge-heather tundra grew in the studied area. A seabird colony existed on Shemya from 4,600 to 2,400 cal year BP according to stable isotope analysis. During a period of at least 2,200 years, nitrogen enrichment led to the development of ornithogenic herb meadows with a high presence of Apiaceae. A long-term increase in δ15N above 9-10‰ led to radical shifts in vegetation. Noticeable reduction of seabird colonies due to human hunting led to grass-meadows spreading. After a prolonged decrease δ15N below 9-10‰ (2,400 cal year BP to present), there was a shift toward less productive sedge-tundra communities. However, the significant enrichment of guano affected only the coastal vegetation and did not alter the inland Shemya Island.


Repetitive sequences in the ITS1 region of ribosomal DNA in congeneric microphallid species (Trematoda: Digenea).
In searching for species-specific DNA sequences of microphallid species (Digenea, Trematoda) we examined the ribosomal internal transcribed spacer regions (ITS) of three closely related species (Levinseniella group) hosted by mud snails (first intermediate host) and marine crustaceans (second intermediate host). In the ITS1 region we found consistent patterns of repeating sequences of 130 bp. Within each main repeat there was a varying number of subrepeats specific for each of the species. All repeats including subrepeats were identified by a similar starting sequence: 5'-CCTGTGG-3'. As this sequence has close resemblance to the chi sequence 5'-GCTGGTGG-3' found in phage lambda we speculate if it serves the same function as a recombination hotspot. Alternatively but less likely, it could be an inactive, mutational relic of a sequence that once served this purpose.


Porcine epidemic diarrhea virus (PEDV) ORF3 protein inhibits cellular type I interferon signaling through down-regulating proteins expression in RLRs-mediated pathway.
Porcine epidemic diarrhea virus (PEDV) is an entero-pathogenic coronavirus, which belongs to the genus Alphacoronavirus in the family Coronaviridae, causing lethal watery diarrhea in piglets. Previous studies have shown that PEDV has developed an antagonistic mechanism by which it evades the antiviral activities of interferon (IFN), such as the sole accessory protein open reading frame 3 (ORF3) being found to inhibit IFN-β promoter activities, but how this mechanism used by PEDV ORF3 inhibits activation of the type I signaling pathway remains not fully understood. Thus, in this present study, we showed that PEDV ORF3 inhibited both polyinosine-polycytidylic acid (poly(I:C))- and IFNα2b-stimulated transcription of IFN-β and interferon-stimulated genes (ISGs) mRNAs. The expression levels of antiviral proteins in the retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs)-mediated pathway was down-regulated in cells with over-expression of PEDV ORF3 protein, but global protein translation remained unchanged and the association of ORF3 with RLRs-related antiviral proteins was not detected, implying that ORF3 only specifically suppressed the expression of these signaling molecules. At the same time, we also found that the PEDV ORF3 protein inhibited interferon regulatory factor 3 (IRF3) phosphorylation and poly(I:C)-induced nuclear translocation of IRF3, which further supported the evidence that type I IFN production was abrogated by PEDV ORF3 through interfering with RLRs signaling. Furthermore, PEDV ORF3 counteracted transcription of IFN-β and ISGs mRNAs, which were triggered by over-expression of signal proteins in the RLRs-mediated pathway. However, to our surprise, PEDV ORF3 initially induced, but subsequently reduced the transcription of IFN-β and ISGs mRNAs to normal levels. Additionally, mRNA transcriptional levels of signaling molecules located at IFN-β upstream were not inhibited, but elevated by PEDV ORF3 protein. Collectively, these results demonstrate that inhibition of type I interferon signaling by PEDV ORF3 can be realized through down-regulating the expression of signal molecules in the RLRs-mediated pathway, but not via inhibiting their mRNAs transcription. This study points to a new mechanism evolved by PEDV through blockage of the RLRs-mediated pathway by ORF3 protein to circumvent the host's antiviral immunity.


Tenascin-cytotactin (TN-C) variants in pseudophakic/aphakic bullous keratopathy corneas.
To examine pseudophakic/aphakic bullous keratopathy (PBK/ABK) human corneas for patterns of expression of tenascincytotactin (TN-C) variants known to mediate specific cellular functions, viz. anti-adhesion (high molecular mass (M(r))) and adhesion (low/intermediate M(r)). PBK/ABK corneas were selected to encompass only those with bullae and without inflammation, scarring or neovascularisation. Serial sections from these and normal corneas were labelled with antibodies BC-4 (recognising all TN-C variants) and BC-2 (specific for the high M(r) TN-C variant). Bound antibody was revealed with an avidin-biotin peroxidase technique. In a given pair of corneal sections, positivity with BC-4 but not BC-2 indicates localisation of low/ intermediate M(r) TN-C variants and absence of the high M(r) TN-C variant. BC-2 identifies the high M(r) variant. There was no immunostaining with either BC-2 or BC-4 in normal corneas except at the corneoscleral interface, where both BC-2 and BC-4 were immunolocalised. In PBK/ABK corneas, BC-2 staining was seen in 5 of 13 corneas and was restricted mainly to epithelial basement membrane (BM) overlying bullae. BC-2 did not label the stroma. BC-4 immunostaining was present in all PBK/ABK corneas and was localised in epithelial BM, both epithelial and stromal borders of bullae, pannus, endothelial BM and in oedematous stromal regions. TN-C variants are differentially expressed in PBK/ABK corneas. The high M(r) variant is restricted mainly to epithelial BM overlying bullae, while low/intermediate M(r) variants occur in epithelial BM, both epithelial and stromal borders of bullae, and in pannus. Given the in vitro functions of TN-C, a role for promoting epithelial dehiscence and reattachment to the substratum in PBK/ABK corneas by high and low/intermediate M(r) variants respectively is likely.


Synthesis and characterization of (1-13C) Phe9 gramicidin A. Effects of side chain variations.
The synthesis of (1-13C)-Phe9-gramicidin (90% enriched) was carried out by the solid phase method. The peptide was removed from the resin by treatment with ethanolamine, deblocked, formylated and purified by preparative t.l.c. to obtain the gramicidin analog in an overall yield of 24%. The peptide was verified and characterized by high pressure liquid chromatography, carbon-13 nuclear magnetic resonance, circular dichroism and single channel currents. Single channel conductances were found to be similar to those of (1-13C)-Phe11-GB but significantly lower than that of gramicidin A. When this gramicidin analog was incubated with phospholipid, the characteristic channel spectrum was not obtained and interaction with sodium ion was not observed. A possible explanation for this behavior is discussed.


Redistribution of local hepatic blood flow during acute bleeding and prolonged hemorrhagic hypotension studied using fluorochromed plasma proteins and surface PO2 measurements.
The present study was performed in order to study the development of sinusoidal blood flow heterogeneity during stepwise bleeding and during prolonged hemorrhagic hypotension. Two methods have been applied: Hepatic surface oxygen tension was measured by a multi-gold-wire-cathode. Sinusoidal perfusion patterns were demonstrated by use of fluorochromed plasma proteins. During stepwise bleeding the relative decrease of the five individual sPO2 curves showed different slopes in the same rat thus indicating a flow dependent heterogeneity. The corresponding perfusion patterns were characterized by an irregular convection front. Time dependent flow heterogeneity resulted in a perfusion pattern showing predominantly perfused sinusoids and low or no flow sinusoids, whereby the predominantly perfused sinusoids might be taken as sinusoidal shunts. It is concluded that stepwise bleeding results in a functional redistribution depending upon the degree of reduced flow, while long time induced microcirculatory changes result in sinusoidal shunt flow depending upon morphological alterations.


Raman spectroscopy for quantifying cholesterol in intact coronary artery wall.
The chemical composition of vascular lesions, an important determinant of plaque progression and rupture, can not presently be determined in vivo. Prior studies have shown that Raman spectroscopy can accurately quantify the amounts of major lipid classes and calcium salts in homogenized coronary artery tissue. This study determines how the relative cholesterol content, which is calculated from Raman spectra collected at the luminal surface of an artery, is related to its depth in an intact arterial wall. Raman spectra of human atherosclerotic plaques were measured after thin tissue layers were successively placed on them. From these spectra, relative cholesterol contents were calculated and used to determine how cholesterol signal strength is attenuated by overlaying tissue. Then, intact artery samples (n = 13) were examined spectroscopically, sectioned and stained specifically for cholesterol. Images of these sections were digitized, and image intensities were related to cholesterol content. These cholesterol amounts were weighed appropriately for depth into the tissue and area-integrated for comparison with spectroscopy results. A decaying exponential curve was fit to the layer study data (r2 = 0.97) and showed that approximately 300 microm of tissue attenuates cholesterol signals by 50%. In intact plaques, the spectroscopically-determined cholesterol amounts correlated strongly and linearly with those determined by digital microscopy (r2 = 0.94). With Raman spectroscopy techniques, the cholesterol content of a lesion can be determined by properly accounting for its depth into an arterial wall. Our results suggest that chemical concentrations in an artery wall could be mapped throughout its thickness, possibly by combining Raman spectroscopy methods with other techniques.


Role of water and steric constraints in the kinetics of cavity-ligand unbinding.
A key factor influencing a drug's efficacy is its residence time in the binding pocket of the host protein. Using atomistic computer simulation to predict this residence time and the associated dissociation process is a desirable but extremely difficult task due to the long timescales involved. This gets further complicated by the presence of biophysical factors such as steric and solvation effects. In this work, we perform molecular dynamics (MD) simulations of the unbinding of a popular prototypical hydrophobic cavity-ligand system using a metadynamics-based approach that allows direct assessment of kinetic pathways and parameters. When constrained to move in an axial manner, the unbinding time is found to be on the order of 4,000 s. In accordance with previous studies, we find that the cavity must pass through a region of sharp wetting transition manifested by sudden and high fluctuations in solvent density. When we remove the steric constraints on ligand, the unbinding happens predominantly by an alternate pathway, where the unbinding becomes 20 times faster, and the sharp wetting transition instead becomes continuous. We validate the unbinding timescales from metadynamics through a Poisson analysis, and by comparison through detailed balance to binding timescale estimates from unbiased MD. This work demonstrates that enhanced sampling can be used to perform explicit solvent MD studies at timescales previously unattainable, to our knowledge, obtaining direct and reliable pictures of the underlying physiochemical factors including free energies and rate constants.


Regional localization of the lactase-phlorizin hydrolase gene, LCT, to chromosome 2q21.
The gene LCT which codes for the intestinal disaccharidase lactase-phlorizin hydrolase has previously been mapped, using somatic cell hybrids and linkage analysis, using the CEPH families, to chromosome 2. We describe here the regional localization of LCT to chromosome 2q21 by polymerase chain reaction (PCR) analysis of somatic cell hybrids and in situ hybridization. LCT is closely linked to D2S44, with a lod score of 30.6 at theta = 0.10.


Developing an openly accessible multi-dimensional small area index of 'Access to Healthy Assets and Hazards' for Great Britain, 2016.
Health geographers have been long concerned with understanding how the accessibility of individuals to certain environmental features may influence health and wellbeing. Such insights are increasingly being adopted by policy makers for designing healthy neighbourhoods. To support and inform decision making, there is a need for small area national level data. This paper details the creation of a suite of open access health indicators, including a novel multidimensional index summarising 14 health-related features of neighbourhoods for Great Britain. We find no association of our overall index with physical health measures, but a significant association to mental wellbeing.


Is there something unique about marriage? The relative impact of marital status, relationship quality, and network social support on ambulatory blood pressure and mental health.
Having close social relationships and being married specifically have been reliably associated with health benefits including lower morbidity and mortality. The purpose of this study was to examine the influence of marital status, relationship quality, and network support on measures of psychological and cardiovascular health. We examined ambulatory blood pressure (ABP) among 204 married and 99 single males and females (N = 303). We found that both marital status and marital quality were important. Married individuals had greater satisfaction with life (SWL) and blood pressure dipping than single individuals. High marital quality was associated with lower ABP, lower stress, less depression, and higher SWL. Importantly, contrasting those who are unmarried with those in low-quality marriages, we find that single individuals had lower ABP-suggesting that single individuals fare better than their unhappily married counterparts. Likewise, having a supportive network did not moderate (i.e., buffer) the effects of being single or unhappily married. Findings indicate being married per se is not universally beneficial, rather, the satisfaction and support associated with such a relationship is important. However, marriage may be distinctive, as evidence further suggests that support from one's network does not compensate for the effect of being single. These results highlight the complexities in understanding the influence of social relationships on long-term health, and they may help clarify the physiological pathways by which such associations exist.


Bilateral ectopic pheochromocytomas: computed tomographic and sonographic correlation.
We describe a case of clinically unsuspected bilateral ectopic pheochromocytomas in a young woman. The bilateral retroperitoneal masses were located in the lower sympathetic ganglia at the level of the organ of Zuckerlandl. Computed tomography (CT) accurately identified the location and vascular characteristics of these neoplasms and provided access for percutaneous biopsy. Sonographic correlation is also presented.


Association between degree of bone-erosion and synovial fluid-levels of tumor necrosis factor alpha in the knee-joints of patients with rheumatoid arthritis.
To determine whether concentrations of cytokines and matrix-degrading enzymes in synovial fluid of patients with rheumatoid arthritis or osteoarthritis are associated with the degree of bone-destruction in the same joint. Determination of Interleukin-1 alpha, IL-1 beta, IL-1-receptor-antagonist, IL-6, IL-8, tumor necrosis factor alpha (by ELISA), collagenase-activity and caseinase-activity (by substrate-assays) in the SF (knee) of patients with RA (n42) or OA (n35). The degree of bone-destruction was assessed radiographically. SF cytokine- and enzyme-levels were higher in patients with RA than in those with OA. In the RA group, SF-levels of TNF alpha were positively correlated with the degree of bone destruction of the respective joint. No correlation was found between radiographically assessed joint changes and SF-concentrations of other cytokines, enzyme activities, serum CRP, or duration of disease. In the OA-group, none of the examined parameters was associated with the degree of joint destruction. Our data may support the assumption of TNF alpha playing an important role in joint destruction in RA. Possible alternative conclusions are discussed.


An exploration of the professional identity of clinical academics using repertory grid technique.
Clinicians who divide their time between clinical work and research have contributed to some of the most fundamental breakthroughs in medicine in recent history, yet their role is not always well-understood or valued. Understanding the factors which contribute to career success for clinical academics is critical for supporting this workforce. Social Cognitive Career Theory (SCCT) provides a conceptual framework for career success, incorporating personal and environmental factors. The aim of this study is to explore clinical academics' construal of successful clinical academic practice and to contribute to a holistic view of the professional identity of the clinical academic. Using a constructivist technique, repertory grid, the authors interviewed ten clinical academics at different career stages in one-to-one structured interviews conducted virtually between November 2020 and April 2021. Data from the interviews were analysed qualitatively and quantitatively. Common themes were identified, analysed, and ranked according to importance with respect to successful clinical academic practice. Using SCCT as a framework, constructs were categorised as personal factors, organisational factors, competencies and person-environment fit. A differential analysis between established/trainee and female/male participants was carried out. One hundred and thirty-three constructs were elicited and categorised into 20 themes (constructs). There was consensus among participants that 6 were of high importance with respect to successful clinical academic practice, 8 of intermediate and 4 of low importance, with no consensus on 2 constructs. Personal factors of high importance include innovation and integrity. Competencies including research and teaching skills are highly important, and ability to collaborate is also considered central to successful clinical academic practice. Female participants expressed greater concerns about the impact of familial responsibilities on career progression. This study highlights the importance of interactions between the person and environment, and characterises the important attributes of successful clinical academics including personal factors such as integrity and innovation.


Therapeutic potential of aptamer-siRNA conjugates for treatment of HIV-1.
Therapeutic strategies designed to treat HIV infection with combinations of antiviral drugs have proven to be the best approach for slowing the progression to AIDS. Despite the great success of highly active antiretroviral therapy (HAART), drug resistance and toxicity issues still remain a concern for some individuals. Therefore, alternative therapeutic strategies need to be developed to overcome these limitations. Nucleic acid-based therapeutics have been considered as an alternative to the currently used antivirals. In this regard, RNA interference (RNAi) can function as a gene-specific therapeutic option for controlling HIV-1 replication. Another type of therapeutic nucleic acid - aptamers - shows promise as a new and potent class of anti-HIV agent and can additionally function as a cell-type-specific delivery vehicle for targeted RNAi. The combined use of small interfering RNA (siRNAs) and aptamers could effectively block viral replication and prevent the emergence of resistant variants. In this review, we recapitulate recent progress and the therapeutic potential of aptamer-siRNA conjugates in the treatment of HIV infection.


Impact of shift work on dementia: a systematic review and dose-response meta-analysis.
Although shift work has been reported as having a link to dementia, evidence remains inconsistent, and a comprehensive dose-response meta-analysis of the association is still lacking. We therefore conducted this meta-analysis to explore the association between shift work and the risk of dementia. Systematic review and dose-response meta-analysis. PubMed, Embase, and Web of Science databases were systematically searched. Fixed or random-effects models were used to estimate the summary relative risks (RRs) and 95% confidence intervals (95% CIs). Generalized least squares regression was used to estimate dose-response associations, and restricted cubic splines were used to examine possible linear or non-linear associations. Five articles (10 studies) with 72,999 participants and 23,067 cases were eventually included in the meta-analysis. The summary RRs and 95% CIs of dementia risk with shift work and night shift work versus daytime work were 1.13 (95% CI: 1.05-1.21, I2 = 46.70%) and 1.13 (95% CI: 1.03-1.24, I2 = 9.20%), respectively. The risk of dementia increased by 1% (RR = 1.01, 95% CI: 1.01-1.02, I2 = 41.3%) with each 1-year increase in the duration of shift work. We found a non-linear dose-response association between the duration of shift work and the risk of dementia (Pnon-linearity = 0.006). Though the shape of the curve was steeper with the duration of shift work <7 years, the increase was more gradual after 7 years. Our findings suggest that shift work may be a risk factor for future dementia and that controlling the length of shift work is a feasible measure that may contribute to prevent dementia.


Demonstration of an autoreceptor modulating the release of [3H]5-hydroxytryptamine from a synaptosomal-rich spinal cord tissue preparation.
A superfusion system employed to measure the K+-stimulated release of [3H]5-hydroxytryptamine [(3H]5-HT, [3H]serotonin) from a synaptosomal-rich spinal cord tissue preparation was carefully characterized, then used to examine the regulation of spinal 5-HT release. Spinal 5-HT release is apparently modulated by an autoreceptor. Exogenous 5-HT depressed, in a concentration-dependent manner, the K+-stimulated release of [3H]5-HT. Similarly, lysergic acid diethylamide (LSD) produced a concentration-dependent decrease in [3H]5-HT release. Methiothepin and quipazine blocked the inhibition of release induced by exogenous 5-HT. The 5-HT2 receptor antagonists spiperone and ketanserin failed to alter the action of 5-HT at the spinal 5-HT autoreceptor. Spiperone and ketanserin were shown, however, to alter the storage of [3H]5-HT. When used in concentrations greater than 10 nM, the drugs evoked increases in basal [3H]5-HT and [3H]5-hydroxyindoleacetic acid ( [3H]5-HIAA) effluxes which were independent of the presence of calcium ions. A good agreement existed between the potencies of drugs for modifying autoreceptor function and their abilities to compete for high-affinity [3H]5-HT binding in the spinal cord (designated 5-HT1). Furthermore quipazine, in concentrations that preferentially interact with the 5-HT1B subtype, antagonized the actions of exogenous 5-HT on K+-stimulated release. Spiperone, in a concentration that approximated the affinity constant of 5-HT1A sites for the drug, was ineffective in altering the ability of exogenous 5-HT to modulate K+-stimulated [3H]5-HT release. These results suggest that 5-HT1B sites are associated with serotonergic autoreceptor function in the spinal cord.


Noradrenaline and avoidance learning in the rat.
The selective neurotoxin 6-hydroxydopamine was used to deplete forebrain noradrenaline to less than 5% of control values and the learning capabilities of the depleted animals examined on a two-way active avoidance task. Noradrenaline depleted animals learned the two-way active avoidance task more quickly than controls and required fewer training trials to reach acquisition criterion. Twenty-four hour retention was not altered by the lesion, but significant resistance to extinction was seen when electric footshock was no longer presented. More detailed analysis of the improved acquisition shown by the lesioned animals revealed that the major effect lay in a reduced freezing response to footshock. This freezing tended to slow down learning in the control animals, since it was incompatible with the required two-way active avoidance response. No alteration was seen in sensory detection thresholds for electric footshock or in spontaneous locomotor activity in the absence of shock. These results are discussed in relation to theories of noradrenaline function in learning and memory and, more recently, in fear and anxiety.


Moving from analogue to digital workflows in dentistry: Understanding undermilling and overmilling as detrimental factors in fabricating CAD/CAM crowns.
Due to the limitations caused by the specific dimensions of milling burs, milling machines may inaccurately fabricate restorations by either over-reducing or under-reducing restorative material, respectively known as "overmilling" and "undermilling". Overmilling occurs when the bur is unable to accommodate areas smaller than the size of the bur, especially at cusp tips and sharp line angles. This results in excess cement space and a weaker restoration. Undermilling occurs when small concavities are ignored by the larger milling bur, particularly at the margins. This results in inadequate seating of the restoration and open margins due to premature binding, which can lead to restorative failure. To prevent these processing errors, clinicians should ensure that all incisal edges and cusp tips are flattened and rounded, verify that there are no undercuts, and prepare teeth with 1mm thick, smooth finish lines. Furthermore, clinicians may use the preparation check and milling simulation step of computer-aided design/computer-aided manufacturing (CAD/CAM) software to confirm the preparation is adequate and the restorative design does not have potential areas that will lead to overmilling.


The heterogeneity of fibrosis and angiogenesis in endometriosis revealed by single-cell RNA-sequencing.
Fibrosis, angiogenesis and chronic inflammation are the inherent characteristics of endometriosis (EMS). The cellular heterogeneity of ectopic and non-ectopic endometrium by single-cell RNA-sequencing (scRNA-seq)at secretory phase without the disturbance of hormone drugs hasn't been explored so far. In this study, scRNA-seq was adopted to explore the properties of ectopic endometrium (ECE), eutopic endometrium (EUE) and normal endometrium (NOE) at secretory phase. We found that (i) The proportion of myofibroblasts, pericytes, endothelial cells and macrophages in ECE overwhelms that of non-ectopic tissues (EUE and NOE), and Myofibro.C2 was the predominant myofibroblast sub-cluster in ECE. (ii) Myofibroblasts were mainly fibroblast-to-myofibroblast transdifferentiation (FMT) and pericytes were endothelial cell-dependent differentiation in ECE. (iii) Both myofibroblasts and pericytes had a low differentiation potential. (iv) The increased inflammation score, deceased NK cells, T cell exhaustion score and antigen-presenting capacity in ECE confirmed the inflammatory properties and immunodeficiency of ECE. These findings suggested that myofibroblasts, pericytes and macrophages may be the potential targets for anti-fibrotic, anti-angiogenic and anti-inflammatory therapy of EMS.


Sexual dimorphism in between and within patch movements of a monophagous insect: Tetraopes (Coleoptera: Cerambycidae).
Movements of milkweed beetles (Tetraopes tetraophthalmus and Tetraopes femoratus) were monitored using mark-release-recapture techniques to compare the within- and between-patch movement patterns of individuals in a naturally occurring population. Within-patch and between-patch movement patterns differ in both frequency and distance traveled for males and females of both study species. Males move farther than females and are more likely to move between patches. Individuals recognize and alter movement upon encountering patch edges. Results suggest that laboratory estimates of Tetraopes vagility give a misleading indication of actual dispersal.


Secondary sensory cells in the gravity receptor system of the statocyst of Octopus vulgaris.
The presence of secondary sensory cells in the Octopus gravity receptor system has been demonstrated. In serial thin sections of the receptor cells (hair cells) no axons were found leaving the cells. Instead, synapses were observed with synaptic vesicles lying inside the receptor cells. Both data clearly indicate that the receptor hair cells represent secondary sensory cells. In addition, efferent contacts to the receptor cells could be confirmed.


Role of endoscopic stenting for biliary strictures in chronic pancreatitis.
To review the published work concerning the role of biliary stenting for chronic pancreatitis-related strictures. A case study in which multiple plastic stents are used to manage a chronic pancreatitis biliary stricture is presented, and the published work reviewed. There has been a gradual evolution in the endoscopic management of distal biliary strictures secondary to chronic pancreatitis. Most early series used single (usually 10 F) plastic stents for varying time periods. Long-term stricture resolution occurred in only approximately 25% of patients and stent-related complications were high if stent exchanges were not performed routinely every 3-4 months. Recent studies using multiple (≥ 3) 10 F stents placed sequentially every few months for approximately 12 months have resulted in resolution of biliary strictures in up to 90% of patients. In general, the use of both uncovered and partially covered self-expandable metal stents for biliary strictures due to chronic pancreatitis have been disappointing due to problems with epithelial hyperplasia involving the uncovered portions of the self-expandable metal stents resulting in late stent occlusion and other problems. Similarly, early published data does not at this stage support the routine use of fully covered self-expandable metal stents because of unacceptable stent-related complications. Chronic pancreatitis-related biliary strictures should be managed initially with sequentially-placed multiple 10 F plastic stents for approximately 12 months.


Frequency shifts due to the interference of resolved peaks in magnitude-mode Fourier-transform ion cyclotron resonance mass spectra.
We have obtained relationships for frequency shifts resulting from the interference of spectral components for the magnitude mode Fourier transform. The approximation of a weak perturbation of well resolved peaks has been used. Both the low- and high-pressure limits for Fourier-transform ion cyclotron resonance (FTICR) operation have been considered. We have found that the shifts can be either negative or positive, depending on the initial phase and/or the choice of the time-domain interval. The magnitude of shifts generally does not exceed the peak width. In the approximation of small perturbations the shifts produced by multiple peaks are additive. We have compared theoretical results with experimental shifts for isotopic clusters of multiply charged insulin. Up to 1 ppm frequency variations were experimentally observed for the insulin 5+ charge state, consistent with theoretical estimates. The peak interference is of particular significance in the case of bio-molecular mass spectra having a large number of peaks and covering a considerable dynamic range (i.e., relative abundance). We conclude that the common mass measurement procedure based on the location of the magnitude mode maxima of well resolved peaks can result in systematic mass measurement errors. The relationships obtained provide corrections for the frequency shifts and thus improve the mass measurement accuracy.


Impaired venous return causes circulatory failure in experimental pancreatitic shock in dogs.
The haemodynamic effects in the early phase of canine acute experimental pancreatitis (AP) were studied using a cardiac catheterization technique. AP was induced in anaesthetized dogs with an infusion of trypsin-sodium-taurocholate into the pancreatic duct. The initial haemodynamic measurements were performed after the preparation of the animal and 5 min after the induction of AP. Thereafter, pressure and volume parameters were measured at 10 min intervals. AP induced significant increases in heart rate, dP/dtmax and mean arterial pressure, but a decrease in Vmax 5 min after the induction of AP. After the initial phase, the heart rate remained significantly increased, while constant and significant decreases of stroke volume, cardiac output, end-diastolic volume and end-diastolic pressure developed. The parameters of the contractility of the left ventricle were not affected to the same extent. It is suggested that the circulatory failure observed in AP, characterised by a prompt reduction of cardiac output, was primarily due to a heavy reduction in preload. This supports the theory that cardiac output is primarily affected by impaired venous return with consequently decreased preload rather than by a loss of ventricular contractility. Hence, the existence of a myocardial depressant factor in the early phase of experimental AP does not gain support from the present results.


The in vitro screening of methylated 4-oxypiperidine compounds for inhibition of protein synthesis in a rabbit reticulocyte cell-free translation system.
A variety of methylated 4-oxypiperidine derivatives were tested for their ability to inhibit protein synthesis in vitro. A direct correlation was found between the extent of methylation of these compounds and their inhibitory activity in a rabbit reticulocyte lysate cell-free translation system.


The Positive Regulation of eNOS Signaling by PPAR Agonists in Cardiovascular Diseases.
Increasing evidence shows that activation of peroxisome proliferator-activated receptors (PPARs) plays an essential role in the regulation of vascular endothelial function through a range of mechanisms, including non-metabolic. Among these, the PPAR-mediated activation of endothelial nitric oxide synthase (eNOS) appears to be of considerable importance. The regulated and sustained bioavailability of nitric oxide (NO) in the endothelium is essential to avoid the development of cardiovascular diseases such as hypertension or atherosclerosis. Therefore, a deeper understanding of the different effects of specific PPAR ligands on NO bioavailability could be useful in the development of novel or multi-targeted PPAR agonists. In this review, we report the most meaningful and up-to-date in vitro and in vivo studies of the regulation of NO production performed by different PPAR agonists. Insights into the molecular mechanisms of PPAR-mediated eNOS activation are also provided. Although findings from animal studies in which the activation of PPARα, PPARβ/δ, or PPARγ have provided clear vasoprotective effects have been promising, several benefits from PPAR agonists are offset by unwanted outcomes. Therefore, new insights could be useful in the development of tissue-targeted PPAR agonists with more tolerable side effects to improve treatment options for cardiovascular diseases.


Two novel mutations of the PAX6 gene causing different phenotype in a cohort of Chinese patients.
Aniridia (AN) is a rare congenital panocular disorder caused by the mutations of the paired box homeotic gene 6(PAX6) gene. The PAX6gene is also involved in other anterior segment malformations including Peters anomaly. We studied the PAX6gene mutations in a cohort of affected individuals with different clinical phenotype including AN, coloboma of iris and choroid, or anterior segment malformations. Six unrelated families and 10 sporadic patients were examined clinically. After informed consent was obtained, genomic DNA was extracted from the venous blood of all participants. Mutation screening of all exons of the PAX6gene was performed by direct sequencing of PCR-amplified DNA fragments. Multiplex ligation-dependent probe amplification (MLPA) was performed to detect large deletions. By clinical examination, the patients and the pedigrees were divided into the following three groups: AN, coloboma of iris and choroids, and the anterior segment malformations including peters anomaly. Sequencing of the PAX6gene, three intragenic mutations including a novel heterozygous splicing-site mutations c.357-3C>G (p.Ser119fsX) were identified in the patients of the AN group. A novel missense mutation c.643T>C (p.S216P) was detected in the anterior segment malformation group. The mutation p.S216P located in the homeodomain region of the PAX6 caused the phenotype of Peters anomaly in family A6 with different expressing. Through MLPA analysis, a large deletion including the whole PAX6gene and DKFZ p686k1684gene was detected in one sporadic patient from the AN group. Neither intragenic mutation nor large deletion was identified in the group with coloboma of iris and choroid. Our findings further confirmed that different kind of mutations might cause different ocular phenotype, and clearly clinical phenotype classification might increase the mutation detection rate of the PAX6gene.


Halogen bonding to the azulene π-system: cocrystal design of pleochroism.
We describe the rational development of a design for pleochroic molecular solids, by using C-Iπ halogen bonds to pre-organise different chromophores in a cocrystal. Using the structure of a known naphthalene cocrystal as a blueprint, we demonstrate how a highly robust C-Iπ motif permits the systematic exchange of original cocrystal components with azobenzene and azulene, resulting in optically interesting dichroic or pleochroic materials.


Inhibition of thymidylate synthase by the diastereoisomers of leucovorin.
The clinical formulation of leucovorin (5-CHO-FH4) is a mixture of diastereoisomers with markedly different pharmacologic properties. Comparatively little information is available concerning the cellular pharmacology of reduced folate stereoisomers, due largely to the difficulty in preparing sufficient quantities of these compounds for in vitro use. Recent improvements in HPLC technology have now facilitated this process, enabling studies of folate stereochemistry on a larger scale. Using purified (6R) and (6S) leucovorin, we examined the effects of these compounds on the enzymatic activity of Lactobacillus casei thymidylate synthase (TS) in a cell-free system. The natural (6S), unnatural (6R), and racemic (6R,S) leucovorin preparations inhibited TS activity by 50% at concentrations of 0.11, 2.1, and 0.52 mM, respectively. Dixon plots demonstrated the inhibition to be competitive, with Ki values of 85 microM, 1.59 mM, and 385 microM for (6S), (6R), and (6R,S) leucovorin, respectively. In view of the high doses of leucovorin given clinically and the slow clearance of the unnatural isomer, our observations suggest that leucovorin may have important direct inhibitory effects on folate-requiring enzymes.


Strategies for specialty training of healthcare professionals in low-resource settings: a systematic review on evidence from stroke care.
The greatest mortality and disability from stroke occurs in low- and middle-income countries. A significant barrier to implementation of best stroke care practices in these settings is limited availability of specialized healthcare training. We conducted a systematic review to determine the most effective methods for the provision of speciality stroke care education for hospital-based healthcare professionals in low-resource settings. We followed the PRISMA guidelines for systematic reviews and searched PubMed, Web of Science and Scopus for original clinical research articles that described or evaluated stroke care education for hospital-based healthcare professionals in low-resource settings. Two reviewers screened titles/abstracts and then full text articles. Three reviewers critically appraised the articles selected for inclusion. A total of 1,182 articles were identified and eight were eligible for inclusion in this review; three were randomized controlled trials, four were non-randomized studies, and one was a descriptive study. Most studies used several approaches to education. A "train-the-trainer" approach to education was found to have the most positive clinical outcomes (lower overall complications, lengths of stay in hospital, and clinical vascular events). When used for quality improvement, the "train-the-trainer" approach increased patient reception of eligible performance measures. When technology was used to provide stroke education there was an increased frequency in diagnosis of stroke and use of antithrombotic treatment, reduced door-to-needle times, and increased support for decision making in medication prescription was reported. Task-shifting workshops for non-neurologists improved knowledge of stroke and patient care. Multidimensional education demonstrated an overall care quality improvement and increased prescriptions for evidence-based therapies, although, there were no significant differences in secondary prevention efforts, stroke reoccurrence or mortality rates. The "train the trainer" approach is likely the most effective strategy for specialist stroke education, while technology is also useful if resources are available to support its development and use. If resources are limited, basic knowledge education should be considered at a minimum and multidimensional training may not be as beneficial. Research into communities of practice, led by those in similar settings, may be helpful to develop educational initiatives with relevance to local contexts.


Kinetic mechanism of 1-N6-etheno-2-aza-ATP hydrolysis by bovine ventricular myosin subfragment 1 and actomyosin subfragment 1. The nucleotide binding steps.
The large change in fluorescence emission of 1-N6-etheno-2-aza-ATP (epsilon-aza-ATP) has been used to investigate the kinetic mechanism of etheno-aza nucleotide binding to bovine cardiac myosin subfragment 1 (myosin-S1) and actomyosin subfragment 1 (actomyosin-S1). The time course of nucleotide fluorescence enhancement observed during epsilon-aza-ATP hydrolysis is qualitatively similar to the time course of tryptophan fluorescence enhancement observed during ATP hydrolysis. In single turnover experiments, the nucleotide fluorescence rapidly increases to a maximum level, then decreases with a rate constant of 0.045 s-1 to a final level, which is about 30% of the maximal enhancement; a similar fluorescence enhancement is obtained by adding epsilon-aza-ADP to cardiac myosin-S1 or actomyosin-S1 under the same conditions (100 mM KCl, 10 mM 4-morpholinepropanesulfonic acid, 5 mM MgCl2, 0.1 mM dithiothreitol, pH 7.0, 15 degrees C). The kinetic data are consistent with a mechanism in which there are two sequential (acto)myosin-S1 nucleotide complexes with enhanced nucleotide fluorescence following epsilon-aza-ATP binding. The apparent second order rate constants of epsilon-aza-ATP binding to cardiac myosin subfragment 1 and actomyosin subfragment 1 are 2-12 times slower than those for ATP. Actin increases the rate of epsilon-aza-ADP dissociation from bovine cardiac myosin-S1 from 1.9 to 110 s-1 at 15 degrees C which can be compared to 0.3 and 65 s-1 for ADP dissociation under similar conditions. Although there are quantitative differences between the rate and equilibrium constants of epsilon-aza- and adenosine nucleotides to cardiac actomyosin-S1 and myosin-S1, the basic features of the nucleotide binding steps of the mechanism are unchanged.


Food Insecurity is Associated With Mortality Among U.S. Adults With Nonalcoholic Fatty Liver Disease and Advanced Fibrosis.
Food insecurity is a growing public health challenge in the United States (U.S.) and has been linked to nonalcoholic fatty liver disease (NAFLD) and advanced fibrosis. However, little is known of how food insecurity impacts mortality risk and health care utilization in chronic liver disease. Using a population-based cohort study of U.S. adults (≥20 years) in the National Health and Nutrition Examination Survey, 1999 to 2014, with NAFLD (estimated by the U.S. Fatty Liver Index) and advanced fibrosis (estimated by the NAFLD fibrosis score, aspartate aminotransferase-to-platelet ratio index, or Fibrosis-4 Index), food security was measured using the Department of Agriculture Food Security Survey Module. The primary outcome was all-cause mortality from National Death Index data and the secondary outcome was health care utilization, defined as ≥2 inpatient and ≥4 outpatient visits, with Cox and logistic regression, respectively, estimating associations between food insecurity and outcomes. Of 34,134 eligible participants (mean age, 47 years; 51% women; 14% in poverty), 4816 had NAFLD and 1654 had advanced fibrosis, with food insecurity present in 28% and 21%, respectively. All-cause age-adjusted mortality was 12 per 1000 person-years among participants with NAFLD (food-secure, 11; food-insecure, 15) and 32 per 1000 person-years among advanced fibrosis participants (food-secure, 28; food-insecure, 50). In multivariable analyses, food insecurity was independently associated with higher mortality among participants with NAFLD (hazard ratio, 1.46; 95% confidence interval [CI], 1.08-1.97) and advanced fibrosis (hazard ratio, 1.37; 95% CI, 1.01-1.86) and greater outpatient health care utilization in participants with NAFLD (odds ratio, 1.32; 95% CI, 1.05-1.67). Food insecurity is significantly associated with greater all-cause mortality in adults with NAFLD and advanced fibrosis. Interventions that address food insecurity among adults with liver disease should be prioritized to improve health outcomes in this population.


Berberine activates PPARδ and promotes gut microbiota-derived butyric acid to suppress hepatocellular carcinoma.
Peroxisome proliferator-activated receptors (PPARs) are a family of ligand-inducible transcription factors that govern various essential metabolic activities in the liver and other organs. Recently, berberine (BBR) has been characterized as a modulator of PPARs; however, the matter of whether PPARs are involved in the inhibitory effect of BBR on hepatocellular carcinoma (HCC) is not well understood. This study aimed to investigate the role of PPARs in the suppressive effect of BBR on HCC and to elucidate the relative mechanism. We studied the role of PPARs in the anti-HCC effects of BBR both in vitro and in vivo. The mechanism whereby BBR regulated PPARs was studied using real-time PCR, immunoblotting, immunostaining, luciferase, and a chromatin immunoprecipitation coupled PCR assay. Additionally, we used adeno-associated virus (AAV)-mediated gene knockdown to address the effect of BBR more effectively. We demonstrated that PPARδ played an active role in the anti-HCC effect of BBR, rather than PPARα or PPARγ. Following a PPARδ-dependent manner, BBR increased BAX, cleaved Caspase 3, and decreased BCL2 expression to trigger apoptotic death, thereby suppressing HCC development both in vitro and in vivo. It was noted that the interactions between PPARδ and the apoptotic pathway resulted from the BBR-induced upregulation of the PPARδ transcriptional function; that is, the BBR-induced activation of PPARδ could mediate the binding with the promoters of apoptotic genes such as Caspase 3, BAX, and BCL2. Moreover, gut microbiota also contributed to the suppressive effect of BBR on HCC. We found that BBR treatment restored the dysregulated gut microbiota induced by the liver tumor burden, and a functional gut microbial metabolite, butyric acid (BA), acted as a messenger in the gut microbiota-liver axis. Unlike BBR, the effects of BA suppressing HCC and activating PPARδ were not potent. However, BA was able to enhance the efficacy of BBR by reducing PPARδ degradation through a mechanism to inhibit the proteasome ubiquitin system. Additionally, we found that the anti-HCC effect of BBR or a combination of BBR and BA was much weaker in mice with AAV-mediated PPARδ knockdown than those in the control mice, suggesting the critical role of PPARδ. In summary, this study is the first to report that a liver-gut microbiota-PPARδ trilogy contributes to the anti-HCC effect of BBR. BBR not only directly activated PPARδ to trigger apoptotic death but also promoted gut microbiota-derived BA production, which could reduce PPARδ degradation to enhance the efficacy of BBR.


Characterization of a sigma class glutathione S-transferase gene in the larvae of the honeybee (Apis cerana cerana) on exposure to mercury.
Glutathione S-transferases (GSTs) are multifunctional enzymes that are mainly involved in detoxification of endogenous and xenobiotic compounds and oxidative stress resistance in insects. In this study, we identified a sigma class GST from Apis cerana cerana (AccGSTs4). The open reading frame of cDNA was 612 bp and encoded a 203 amino acid polypeptide, which exhibited the structural motif and domain organization characteristic of GST. Homology and evolutionary analysis indicated that the induced amino acid sequence of AccGSTs4 belonged to an insect sigma class group. Expression analysis indicated that AccGSTs4 was presented in all stages of development with high level in 4th instar larvae. Immunolocalization further revealed the distribution of AccGSTs4 in 4th instar larvae. RT-qPCR showed that the transcripts of AccGSTs4 from the larvae were upregulated under dietary HgCl(2). The GST activity under stress was higher than the controls fed on HgCl(2)-free diet. Disc diffusion assay provided evidence of recAccGSTs4 resistance to long-term exposure of HgCl(2) stress. Additionally, analysis of 5'-flanking region further clarified the probable expression patterns of AccGSTs4. Taken together, our findings indicate that the larvae AccGSTs4 may play a role in mercury stress response, and it will help to protect honeybees from heavy metals.


Impact of the COVID-19 pandemic on family carers of those with profound and multiple intellectual disabilities: perspectives from UK and Irish Non-Governmental Organisations.
Family carers of people with profound and multiple intellectual disabilities (PMID) experienced a reduction in healthcare services due to the COVID-19 pandemic. Many subsequently turned to Non-Governmental Organisations who worked to support families. However, little research has sought to capture the experiences of family carers or identify effective interventions which might support them. To address these concerns we explored the views of Non-Governmental sector workers across the UK and Ireland who supported families people with PMID during the COVID-19 pandemic. We also sought to explore their views on the characteristics of online support programmes for family carers. METHODS: This study employed a qualitative design using focus groups with participants (n = 24) from five Non-Governmental Organisations across the UK and Ireland. A focus group guide included questions on challenges, supports, coping and resources which helped during lockdown restrictions. Focus groups were held online, were audio recorded and transcribed verbatim. The resulting transcripts were pseudonymised and subjected to thematic analysis. Four themes were identified (i) 'mental and emotional health', (ii) 'they who shout the loudest' (fighting for services), (iii) 'lack of trust in statutory services' and (iv) 'creating an online support programme'. Mental and emotional health emerged as the most prominent theme and included three subthemes named as 'isolation', 'fear of COVID-19' and 'the exhaustion of caring'. The COVID-19 pandemic has increased the vulnerability of family carers who were already experiencing difficulties in accessing services and supports for their families. While Non-Governmental Organisations have been a crucial lifeline there is urgent need to design services, including online support programmes, in partnership with family carers which adequately address their needs.


Decreased IL-1 RA concentration in ascites is associated with a significant improvement in overall survival in ovarian cancer.
Cytokines play a major role in promoting the growth and metastatic spread of cancer cells. Interleukin-1 alpha and beta (IL-1) and IL-1 RA are known to be critically involved in carcinogenesis and in various solid tumors. There are limited data on expression of IL-1 alpha, beta and RA in serum and ascites in patients with advanced ovarian cancer. Objectives of this study were to investigate the level of IL-1 alpha, IL-1 beta and IL-1 RA in serum and ascites from patients with ovarian cancer and their impact on the prognosis. Fifty-three women with ovarian cancer (OC) (33 patients with primary OC and 20 with relapsed OC) and 50 women with benign gynaecological diseases as a control group (CG) were enrolled onto this prospective study. IL-1 alpha, beta and RA levels were analyzed in serum and ascites by ELISA technique. The median age was 55 years (range 19-80) in the ovarian cancer group and 40 years (range 15-89) in the controls. The distribution of histological type of ovarian cancer was as follows: serous-papillary 43 (81.1%), 4 (7.5%) mucinous, 3 (5.7%) endometroid and 3 (5.7%) clear cell carcinoma. The concentrations of IL-1 beta and RA in ascites or peritoneal fluid were significantly increased in patients with OC in comparison to the CG, for both cytokines (p<0.0001); also the concentration of IL-1 RA in serum was increased in OC (p=0.003) vs. CG. An increased level of IL-1 beta in ascites correlated significantly with a poorer histopathological grading (p=0.038). IL-1 RA concentration in ascites was correlated with advanced FIGO stage (p=0.049) and the IL-1 RA serum level with ascites volume (< or =500 ml vs. >500 ml) (p=0.046). Patients with IL-1 RA level in ascites lower than the cut off value of 695.6 pg/ml showed a significant better progression-free median survival (24.6 vs. 12.8 months, p=0.008) and postoperative median overall survival (34.6 vs. 17 months, p=0.01) in comparison to patients with an IL-1 RA level in ascites higher than the cut off level. Additionally, a higher expression of IL-1 beta in serum (p=0.004) and ascites (p=0.05) reduced significantly the progression-free survival. In the multivariate analysis, expression of IL-1 RA in ascites was an independent prognostic factor for good progression-free and postoperative overall survival (HR, 0.39 95% CI, 0.18-0.83, p=0.01, HR, 0.36 95% CI, 0.16-0.8, p=0.01). IL-1 RA levels in ascites lower than the cut off value of 695.6 pg/ml are associated with a significant improvement in postoperative and progression-free survival. IL-1 RA shows a prognostic relevance in ovarian cancer.


Cloning and characterization of the Bacillus sphaericus genes controlling the bioconversion of pimelate into dethiobiotin.
Using 8.8 kb of genetic information from Bacillus sphaericus, it was possible to confer to Escherichia coli bio- strains, including delta bioA-D, bioC-, bioH-, the ability to convert exogenous pimelate into biotin. The bio genes were borne on two recombinant plasmids with inserts of 4.3 kb and 4.5 kb, which had been isolated from a genomic bank of HindIII-digested B. sphaericus DNA, by phenotypic complementation of various E. coli bio mutants. The B. sphaericus bioD and bioA genes were unambiguously identified within the 4.3-kb insert and shown to be closely linked to bioY (coding for a protein with a presently unknown function) and to bioB [Ohsawa et al., Gene 80 (1989) 39-48]. These genes are clustered in the order bioDAYB. The 4.5-kb fragment contains genetic information for three different proteins, the products of bioX, bioW and bioF. Complementation studies using an E. coli bioF mutant and a B. subtilis bio112TG3 strain, revealed that the third ORF of this cluster encodes 7-keto-8-aminopelargonic acid synthetase. A combination of bioW and bioF allows an efficient complementation of E. coli bioC and bioH mutants, provided that pimelate is added to the biotin-depleted growth medium. No function could be identified for the product of bioX. The gene order of this cluster is bioXWF. By sequence analysis, the two cloned DNA fragments were shown to bear overlapping open reading frames and secondary structures at their 3' ends, typical of transcription terminators.(ABSTRACT TRUNCATED AT 250 WORDS)


Glycovaccinology: The design and engineering of carbohydrate-based vaccine components.
Vaccines remain one of the most important pillars in preventative medicine, providing protection against a wide array of diseases by inducing humoral and/or cellular immunity. Of the many possible candidate antigens for subunit vaccine development, carbohydrates are particularly appealing because of their ubiquitous presence on the surface of all living cells, viruses, and parasites as well as their known interactions with both innate and adaptive immune cells. Indeed, several licensed vaccines leverage bacterial cell-surface carbohydrates as antigens for inducing antigen-specific plasma cells secreting protective antibodies and the development of memory T and B cells. Carbohydrates have also garnered attention in other aspects of vaccine development, for example, as adjuvants that enhance the immune response by either activating innate immune responses or targeting specific immune cells. Additionally, carbohydrates can function as immunomodulators that dampen undesired humoral immune responses to entire protein antigens or specific, conserved regions on antigenic proteins. In this review, we highlight how the interplay between carbohydrates and the adaptive and innate arms of the immune response is guiding the development of glycans as vaccine components that act as antigens, adjuvants, and immunomodulators. We also discuss how advances in the field of synthetic glycobiology are enabling the design, engineering, and production of this new generation of carbohydrate-containing vaccine formulations with the potential to prevent infectious diseases, malignancies, and complex immune disorders.


Unusual iodine-131 postablation whole-body scintigraphy patterns in patients after robot-assisted/endoscopic thyroidectomy: Case series.
Robot-assisted/endoscopic thyroidectomy causes irritation due to instruments or implantation of thyroid tissue on the anterior chest wall and lower neck. We present three patients who exhibited unexpected focal lesions on postablation Iodine-131 single-photon emission computed tomography/computed tomography without biochemical and structural diseases. Meticulous surgical techniques are important to prevent complications.


Post-nursing early life macronutrient balance promotes persistent and malleable biometric and metabolic traits in mice.
It is known that dietary factors within the gestational and nursing period affect early life and stably affect later life traits in animals. However, there is very little understanding of whether dietary factors within the early life period from post-nursing to adulthood affect traits in adulthood. To address this, we conducted studies on male C57Bl/6J mice fed from 3 weeks (immediately post-nursing) until 12 weeks (full maturity) using nine different diets varying in all three major macronutrients to parse out the effects of individual macronutrients. Early life macronutrient balance affected body composition and glucose homeostasis in early adulthood, with dietary protein and fat showing major effects. Despite this, mice showed rapid reversal of the effects on body composition and glucose homeostasis of early life diet feeding, upon standard diet feeding in adulthood. However, some traits were persistent, with early life low dietary protein levels stably affecting lean and muscle mass, and early life dietary fat levels stably affecting serum and liver triglyceride levels. In summary, macronutrient balance in the post-nursing early life period does not stably affect adiposity or glucose homeostasis but does impact muscle mass and lipid homeostasis in adulthood, with prominent effects of both protein and fat levels. KEY POINTS: Early life dietary low protein and high fat levels lowered and heightened body mass, respectively. These effects did not substantially persist into adulthood with rapid catch-up growth on a normal diet. Early life protein (negative) and fat (positive) levels affected fat mass. Early life low protein levels negatively affected lean mass. Low protein effects on lower lean and muscle mass persisted into adulthood. Early life macronutrient balance effects did not affect later life glucose homeostasis but early life high fat level affected later life dyslipidaemia. Effects of dietary carbohydrate levels in early and later life were minor.


Sickle cell disease: loss of the blood's WD40?
Sickle cell disease (SCD) is characterized by malformed erythrocytes and results in many vascular complications, including the lysis of a minor proportion of these cells, liberating free hemoglobin, which is a potent scavenger of nitric oxide (NO). SCD involves inflammatory activation, including the upregulation of vascular coagulation. Because NO possesses important anti-coagulant and anti-adhesion properties, the increased scavenging of NO in SCD undoubtedly is a major contributor to the pathology of this disease.


Activation of mouse sperm phosphatidylinositol-4,5 bisphosphate-phospholipase C by zona pellucida is modulated by tyrosine phosphorylation.
Many cellular responses to the occupancy of membrane receptors include the hydrolysis of phosphatidylinositol-4,5 bisphosphate (PIP2) by phospholipase C (PLC) and the subsequent generation of inositol 1,4,5-triphosphate (IP3) and diacylglycerol (DAG). In the gamete interaction system, sperm respond to binding to the egg's extracellular matrix, the zona pellucida (zp), by exocytosis of the acrosome in a process known as the acrosome reaction (AR). Under physiological conditions, zp binding stimulates ARs only after sperm have undergone a final maturation phase, known as capacitation. One of the zp glycoproteins, ZP3, serves as the ligand for sperm plasma membrane receptors and as the trigger for this regulated exocytosis. Both phosphoinositide-linked and tyrosine kinase-mediated pathways participate in the signalling cascade triggered by sperm-zp interaction. This paper reports that stimulation with solubilized zp increased PIP2-PLC enzymatic activity from mouse sperm. ZP3 is the zp component responsible for this stimulation. The effect was abolished by tyrphostin, suggesting that zp activation of PLC was mediated by tyrosine phosphorylation and that gamma was the PLC isoform involved. We show the presence and distribution of PLC gamma 1 in mouse sperm. Immunostaining studies indicate that PLC gamma 1 is restricted to the sperm head. Sperm capacitation induced translocation of PLC gamma 1 from the soluble to the particulate fraction. These data suggest that PLC gamma 1 constitutes a component in the cascade that couples sperm binding to the egg's extracellular matrix with acrosomal exocytosis, a regulated secretory response upon which fertilization depends absolutely.


Life After Retirement: Exploring the Survival Strategies of SSNIT Pensioners in the Greater Accra Region of Ghana.
Ghana's elderly population represents about 6.7% of the national populace. They comprise persons aged sixty years and above, including retirees. The main source of income for some of these individuals, particularly those who worked in the formal sector is the monthly pension they receive as compensation for contributing to Ghana's public pension scheme known as the Social Security and National Insurance Trust (SSNIT) while they were working. However, some beneficiaries of the scheme have raised concerns about the inadequacy of the pension income. Old age is accompanied by other social, economic, and health issues that tend to be detrimental to one's well-being. In this study, the survival strategies adopted by SSNIT pensioners to assuage the challenges they encounter, mainly due to inadequate pensions were explored. The mixed-methods research design was employed to gather data from 437 respondents. Whereas the quantitative findings were analyzed via the Stata software, the framework approach was adopted to analyze the qualitative data. The majority (78.7%) of the pensioners indicated that their pensions were insufficient to meet their needs. In their quest to address the challenges confronting them, the pensioners either adopted survival strategies they considered suitable or resorted to other non-prioritized coping mechanisms perceived to be relevant only in times of urgent necessity. Based on the findings of the study, some recommendations for policy and practice were presented.


Use of Fourier transform infrared spectra of crude bacterial lipopolysaccharides and chemometrics for differentiation of Salmonella enterica serotypes.
To evaluate Fourier transform infrared spectroscopy (FTIR) and chemometrics for differentiating intact cells and crude lipopolysaccharide (LPS) extracts from Salmonella serotypes. Intact cells and crude LPS extracts from six different Salmonella enterica serotypes (Typhimurium, Enteritidis, Thomasville, Brandenburg, Hadar and Seftenberg) were used. The crude Salmonella LPS extracts were visualized using deoxycholic acid-polyacrylamide gel electrophoresis (DOC-PAGE) and appeared heterogeneous on the gel with two exceptions: S. Enteritidis and S. Brandenburg, and S. Thomasville and S. Seftenberg. Canonical variate analysis (CVA) of spectra of crude LPS extracts provided 100% correct classification. CVA of spectra of intact cells was not useful for classifying the Salmonella serotypes, having only 47 and 50% correct classifications in the 1200-900 and 4000-700 cm(-1) regions respectively. These data were confirmed by greater Mahalanobis distances between crude LPS spectra than intact cell spectra. CVA of FTIR spectra of crude LPS extracts from Salmonella serotypes provided a 100% correct serotype classification. This study suggests that the FTIR analytical procedure provides chemical detail as well as a better separation of Salmonella serotypes using spectra of crude LPS extracts than analysis using DOC-PAGE.


Analysis of isolates within species of anuran trypanosomes using random amplified polymorphic DNA.
A total of 20 decamer primers were used to generate random applied polymorphic DNA (RAPD) markers from 5 isolates of Trypanosoma fallisi, 3 isolates of T. ranarum, 2 isolates of T. rotatorium, and 2 isolates of T. rotatorium-like trypanosomes in addition to 2 species from the American Type Culture Collection, T. chattoni (ATCC 50294) and Trypanosoma sp. (ATCC 50295). A slight polymorphism was observed among the four isolates of T. fallisi obtained form American toads, Bufo americanus, collected in Algonquin Park, Ontario, Canada, and an isolate obtained from the same species of host collected in Marquette, Michigan, United States, and produced similarity coefficients ranging from 80.7% to 96.9%. Pronounced polymorphism was recorded among the three isolates of T. ranarum from bullfrogs, Rana catesbeiana, collected in Ontario, Canada, and in Maryland, United States, and from a Northern leopard frog, R. pipiens, collected in Minnesota (USA). The similarity coefficients ranged from 54.7% to 59.5%, suggesting that alleles of these isolates were conserved over a wide geographic range. The high degree of polymorphism observed in two isolates of T. rotatorium from a bullfrog collected in Ontario and two isolates of a T. rotatorium-like parasite from the green frog R. clamitans, collected in Louisiana (USA) suggests that they are different species. These results reflect the high similarity among isolates from the same geographic location and the pronounced polymorphism apparent among isolates from distant geographic locations.


Microbiological risk assessment in Europe: the next decade.
Under the Agreement on the Application of Sanitary and Phytosanitary Measures that came into force in 1995, it was stipulated that all World Trade Organisation (WTO) members are required to ensure that their sanitary and phytosanitary measures are based on assessment of the risks to human, animal or plant life or health. In doing so, they must take into account risk assessment techniques developed by relevant international organisations. WHO and FAO are two such international organisations that are of very high standard and influence. Their executive body, the Codex Alimentarius Commission has developed the concept of Microbiological Risk Assessment (MRA) in a wider framework called Risk Analysis. The aim of Risk Analysis is to provide a global standard for the interpretation of the acceptability of risks associated to foods to which consumers might be exposed. Microbiological Risk Assessment is an essential element of Risk Analysis because it specifies risks related to pathogenic micro-organisms in the food chain on the basis of sound science, combining qualitative and quantitative data in the areas of epidemiology and pathogenicity of micro-organisms with food production and handling. The concept is still in its infancy but is gaining wide acceptance globally. This paper will describe an inventory of MRA developments in Europe carried out under auspicion of the European Commission. It also gives the results of a literature survey concerning scientific publications on the topic MRA in which the activities in Europe are compared to those elsewhere.


Management of adenocarcinoma extending to the middle and upper part of the esophagus.
Of 30 patients with adenocarcinoma of the gastroesophageal junction treated between 1977 and 1982, 11 had tumor spread to the middle or upper thirds of the esophagus. Ten were treated with esophagogastrectomy by the Lewis approach, with two operative deaths. All of the tumors had spread to the periesophageal tissues. The eight surviving patients had relief of dysphagia. Five died with recurrent tumor after a mean survival time of 11 months; three are alive at five and 16 months (without evidence of disease) and 13 months (with recurrence). Adjuvant preoperative radiation therapy was used in four patients, three of whom received concomitant total parenteral nutrition. Despite the poor prognosis of this particularly unfavorable subset of patients with tumors of the gastroesophageal junction, operative resection provides acceptable palliation. Further study of adjuvant modalities is needed.


Contemporary depositional environments of the Omo delta.
Geomorphological and sedimentological studies of depositional environments of the modern Omo River delta and floodplain are essential to an understanding of the Pliocene to Pleistocene Mursi, Nkalabong and Kibish Formations of the Lower Omo Basin (southwestern Ethiopia).


The genetics of the target tissue in rheumatoid arthritis.
The genetic mechanisms that are complementary in predisposing and then establishing disease are yet to be fully elucidated. During a lifetime, the genetic composition of the host is not only hereditary but undergoes rearrangements, integrations, and more subtle single-base pair alterations. These changes can be inconsequential or lead to aberrant and deleterious pathologic changes. In a complex multifactorial disease such as RA, the relative roles of the dynamic versus germline elements of the disease have yet to be fully determined. Further studies of large populations are likely to segregate out factors affecting specific ethnic, clinical, and genetic subgroups.


Induction and modification of erythropoiesis in a suspension culture system derived from long-term bone marrow culture of Syrian hamster.
Cells in a subculture system derived from long-term hamster bone marrow culture were induced to differentiate into hemoglobin-producing cells in response to added erythropoietin. Treatment of the subculture with dimethyl sulfoxide or other 'Friend inducers' but not 5-azacytidine effected no remarkable induction, though they enhanced the effect of externally added erythropoietin. 3-Aminobenzamide, a weak inducer of Friend cell differentiation [Morioka, K., K. Tanaka, M. Ishizawa and T. Ono: Dev. Growth Differ. 24, 507-512 (1982)], had a strong inhibitory effect on erythropoiesis in the subculture system. Phorbol esters and glucocorticoids (both are known to be strong inhibitors of Friend cell differentiation) acted as strong inhibitors of the differentiation in our system as well. The present system was shown to provide a useful in vitro model to compare the differentiation of normal erythroid progenitors with the previously established systems of neoplastic cells.


Deep learning-based coronary computed tomography analysis to predict functionally significant coronary artery stenosis.
Fractional flow reserve derived from coronary CT (FFR-CT) is a noninvasive physiological technique that has shown a good correlation with invasive FFR. However, the use of FFR-CT is restricted by strict application standards, and the diagnostic accuracy of FFR-CT analysis may potentially be decreased by severely calcified coronary arteries because of blooming and beam hardening artifacts. The aim of this study was to evaluate the utility of deep learning (DL)-based coronary computed tomography (CT) data analysis in predicting invasive fractional flow reserve (FFR), especially in cases with severely calcified coronary arteries. We analyzed 184 consecutive cases (241 coronary arteries) which underwent coronary CT and invasive coronary angiography, including invasive FFR, within a three-month period. Mean coronary artery calcium scores were 963 ± 1226. We evaluated and compared the vessel-based diagnostic accuracy of our proposed DL model and a visual assessment to evaluate functionally significant coronary artery stenosis (invasive FFR < 0.80). A deep neural network was trained with consecutive short axial images of coronary arteries on coronary CT. Ninety-one coronary arteries of 89 cases (48%) had FFR-positive functionally significant stenosis. On receiver operating characteristics (ROC) analysis to predict FFR-positive stenosis using the trained DL model, average area under the curve (AUC) of the ROC curve was 0.756, which was superior to the AUC of visual assessment of significant (≥ 70%) coronary artery stenosis on CT (0.574, P = 0.011). The sensitivity, specificity, positive and negative predictive value (PPV and NPV), and accuracy of the DL model and visual assessment for detecting FFR-positive stenosis were 82 and 36%, 68 and 78%, 59 and 48%, 87 and 69%, and 73 and 63%, respectively. Sensitivity and NPV for the prediction of FFR-positive stenosis were significantly higher with our DL model than visual assessment (P = 0.0004, and P = 0.024). DL-based coronary CT data analysis has a higher diagnostic accuracy for functionally significant coronary artery stenosis than visual assessment.


Detection of Angiotensin II type I-receptor antibodies in transplant glomerulopathy.
Transplant glomerulopathy (TG) is an important cause of late graft loss. The role of angiotensin type 1-receptor antibodies (AT1 R-Ab) in TG is not known. All the TG cases (N = 137) between January 2007 and December 2014 (N = 1410) were analyzed. Donor-specific anti-HLA antibodies (DSA) at the time of biopsy and AT1 R-Ab IgG (positive, >17 UI/mL; "at risk," 10-17 UI/mL; negative, <10 UI/mL) in pre-transplant sera (PT-Ab) and at biopsy time (BT-Ab) were studied. AT1 R-PT-Ab+ and AT1 R-BT-Ab+ patients were 16.5% (51.5% "at risk") and 11.5% (27.4% "at risk"), respectively. Clinical correlations were found between AT1 R-Ab and HCV infection, number of transplants, and age. Considering Banff scores, ptc was higher in DSA+ patients vs AT1 R-PT-Ab+ (P = 0.002) or AT1 R-BT-Ab+ (P = 0.001) without differences in g and chronicity score (ci + ct); cg showed lower scores in DSA+ patients vs AT1 R-BT-Ab+ (P = 0.001). Graft survival was not influenced by the presence of AT1 R-Ab, AT1-R-Ab titer or MFI, but we observed a longer graft survival in patients with both AT1 R-BT-Ab+ or "at risk" and DSA+ vs patients positive only for DSA (P = 0.02), for AT1 R-BT-Ab (P = 0.019) or AT1 R-BT-Ab "at risk" (P = 0.039). AT1 R-Ab showed no independent prognostic role in TG in this pilot analysis.


Impact of phenotype on treatment and prognosis of laryngeal malignancies.
An overview of the impact of the phenotype on treatment and prognosis of different laryngeal malignancies is presented.


Development, validity and reliability of the Turkish version of the Hung Postpartum Stress Scale.
The aim of the current study was to translate the Hung Postpartum Stress Scale into Turkish and test the reliability and validity of the Turkish version of the scale. The translation process of the Hung Postpartum Stress Scale into Turkish and the testing of reliability and validity of the newly developed scale on postpartum stress in women are described. The questionnaire was translated using a back-translation technique. Expert review of internal consistency reliability, content validity, factor analysis of construct validity and criterion-related validity were examined. Cronbach's alpha for the Turkish version of the Hung Postpartum Stress Scale was 0.931, indicating very good reliability. Factor analyses resulted in a two factor scale structure: maternal concerns and acceptability of the newborn by the family. Analyses also indicated good test-re-test reliability for the Turkish version of the Hung Postpartum Stress Scale (P < 0.01). The present study describes the design of a Turkish version of the Hung Postpartum Stress Scale. The newly developed scale proved to be reliable and valid and will be a valuable instrument for women's healthcare professionals.


Traumatic brain injury among two-wheeled motor vehicle riders in Utsunomiya, Japan: a comparison between mopeds and motorcycles.
Two-wheeled motor vehicles are dichotomized into mopeds and motorcycles (MCs) based on their engine capacity. While efforts have been made, mostly by European researchers, to evaluate the possible difference in the frequency/severity of riders' traumatic brain injury (TBI) between the two categories, the results have been inconsistent. To evaluate whether such differences exist in Japan, a single-center, retrospective observational study was conducted on two-wheeled motor vehicle riders wearing a helmet during high-energy road traffic accident (RTA). Between January 2011 and December 2017, 127 moped and 128 MC helmeted riders involved in RTAs had been treated in our institution. Moped riders were significantly older than MC riders (50.7 ± 25.0 years vs. 35.1 ± 18.6 years, p = 0.04). The frequency of TBI was significantly higher in moped riders than in MC riders (21.3% vs. 10.9%; p = 0.03). However, the frequency of other bodily injuries did not differ significantly. Among the riders with TBI, neither injury severity nor outcomes differed significantly. Multivariate regression analysis showed that the presence of altered mental status was predictive of TBI (OR 23.398; 95% CI 9.187-59.586; p < 0.001). Moped riders trended to have a higher likelihood of sustaining TBI (OR 2.122; 95% CI 0.870-5.178; p = 0.098). Mopeds are frequently utilized by elderly in Japan, which may have been causally associated with the higher frequency of TBI in moped riders, while other causes, including the difference in helmet type, may also be involved. This study is limited by its small sample size and retrospective design, and multi-center prospective studies are warranted.


Assessment of in vivo organ-uptake and in silico prediction of CYP mediated metabolism of DA-Phen, a new dopaminergic agent.
The drug development process strives to predict metabolic fate of a drug candidate, together with its uptake in major organs, whether they act as target, deposit or metabolism sites, to the aim of establish a relationship between the pharmacodynamics and the pharmacokinetics and highlight the potential toxicity of the drug candidate. The present study was aimed at evaluating the in vivo uptake of 2-Amino-N-[2-(3,4-dihydroxy-phenyl)-ethyl]-3-phenyl-propionamide (DA-Phen) - a new dopaminergic neurotransmission modulator, in target and non-target organs of animal subjects and integrating these data with SMARTCyp results, an in silico method that predicts the sites of cytochrome P450-mediated metabolism of drug-like molecules. Wistar rats, subjected to two different behavioural studies in which DA-Phen was intraperitoneally administrated at a dose equal to 0.03mmol/kg, were sacrificed after the experimental protocols and their major organs were analysed to quantify the drug uptake. The data obtained were integrated with in silico prediction of potential metabolites of DA-Phen using the SmartCYP predictive tool. DA-Phen reached quantitatively the Central Nervous System and the results showed that the amide bond of the DA-Phen is scarcely hydrolysed as it was found intact in analyzed organs. As a consequence, it is possible to assume that DA-Phen acts as dopaminergic modulator per se and not as a Dopamine prodrug, thus avoiding peripheral release and toxic side effects due to the endogenous neurotransmitter. Furthermore the identification of potential metabolites related to biotransformation of the drug candidate leads to a more careful evaluation of the appropriate route of administration for future intended therapeutic aims and potential translation into clinical studies.


Second-Line Treatment of Her2-Positive Metastatic Breast Cancer: Trastuzumab beyond Progression or Lapatinib? A Population Based Cohort Study.
The relative efficacy of lapatinib vs. continuing trastuzumab beyond progression (TBP) in HER2-positive metastatic breast cancer (MBC) patients, who progressed on first-line trastuzumab, is still unclear. The objective of this population based cohort study was to compare outcomes of lapatinib vs. TBP in daily practice. All HER2-positive MBC patients who began second-line anti HER2 therapy between 1st January 2010 and 30th August 2013 were selected from Clalit Health Services' (CHS) electronic database. Available data on patient and disease characteristics and treatments were analyzed. The primary endpoint was overall survival (OS). Outcomes were compared using the Kaplan-Meier (log-rank) method and Cox proportional hazards model. 64 patients received second-line lapatinib and 93 TBP. The two treatment groups were similar in age and co-morbidity rates, but differed in proportion of prior adjuvant trastuzumab (lapatinib: 29.7%, TBP: 16.1%, P = 0.043) and rates of prior brain metastases (lapatinib: 32.8%, TBP: 10.8%, P = 0.01). Lapatinib median OS was 13.0 months (95% CI: 9.5-16.5) vs. 31.0 for TBP (95% CI: 20.6-41.4), P<0.001. On multivariate analysis, longer OS was preserved for TBP, after controlling for differences in age, adjuvant trastuzumab, duration of first-line trastuzumab therapy, brain metastases, visceral metastases and hormonal treatment [Hazard Ratio (HR) = 0.63, 95% CI: 0.40-0.99, P = 0.045]. In this comparative cohort study, OS of HER2-positive MBC patients treated with TBP was significantly longer than with lapatinib. These results might be especially relevant in settings where ado-trastuzumab-emtansine (TDM-1), the current preferred agent in this setting, is not available yet for patients.


Evaluation of the "Kosish Cocktail" in Treating Severe Pain in "Home Care" in Morphine-naïve Communities.
Inavailability of morphine continues to plague most parts of India. Good palliative care must, however, focus on resources that are locally available, culturally acceptable, financially affordable, and easily applicable. These factors were all integral to the development of the "Kosish cocktail" for use in severe pain. This cocktail is a mixture of ketamine, midazolam, pentazocine lactate, and other adjuvants for use in the domiciliary set-up as intermittent subcutaneous injections in a morphine-naïve community. Our aims and objectives were: (1) To assess the efficacy of the "Kosish cocktail" in treating severe pain in terminally ill patients; (2) To assess the safety profile and note any adverse effects; (3) To evaluate its use in domiciliary set-ups in terms of safety and efficacy; (4) To empower the patient and the family in the process of patient care. Eight patients with advanced cancer and severe pain, who were already on WHO Step II drug therapy, were enrolled for this study. The cocktail was administered subcutaneously in every four hours and SOS. Subjective and objective parameters were recorded and the data analyzed using Student's t-test with a P<0.05 being considered significant. There was a statistically significant improvement in the subjective parameters 12 hours after the initiation of therapy, except for the persistence of fatigue. On the basis of this qualitative study, the authors confirm the efficacy and safety of the use of the Kosish cocktail in treating severe pain, and strongly recommend it for newly started hospices, especially for use in the domiciliary set-up.


The unit structure of the locust compound eye.
1. The ommatidium or functional unit of the locust compound eye comprises a compound corneal lens, 4 cone cells, 2 primary pigment cells, 16 secondary pigment cells and 8 retinula cells. 2. All retinula cells run the entire length from the cone to the basal lamina, although two, called the proximal cells, only contribute to the lowest third of the rhabdom, and one of either cell 6 or cell 7 on our arbitrary numbering system forms its axon one third the way up the ommatidium. 3. 84% of the 417 ommatidia examined had five cone cell processes. The position of three cone cell processes (cone threads) is almost invariable with respect to numbered retinula cells but the remaining threads can take any of three intercellular locations. 4. The position of these threads correlates with the number of the cell distally displaced from the rhabdom. We suggest that cone thread position in the developing ommatidium determines some features of retinula cells and we propose a simple model to account for this.


Effects of step length, age, and fall history on hip and knee kinetics and knee co-contraction during the maximum step length test.
Maximum step length is a brief clinical test involving stepping out and back as far as possible with the arms folded across the chest. This test has been shown to predict fall risk, but the biomechanics of this test are not fully understood. Knee and hip kinetics (moments and powers) are greater for longer steps and for younger subjects, but younger subjects also step farther. To separate the effects of step length, age, and fall history on joint kinetics; healthy younger (age=27(5), N=14), older non-fallers (age=72(5), N=14), and older fallers (age=75(6), N=11) all stepped to the same relative target distances of 20-80% of their height. Knee and hip kinetics and knee co-contraction were calculated. Hip and knee kinetics and knee co-contraction all increased with step length, but older non-fallers and fallers utilized greater stepping hip and less stepping knee extensor kinetics. Fallers had greater stepping knee co-contraction than non-fallers. Stance knee co-contraction of non-fallers was similar to young for shorter steps and similar to fallers for longer steps. Age had minimal effects and fall history had no effects on joint kinetics of steps to similar distances. Effects of age and fall history on knee co-contraction may contribute to age-related kinetic differences and shorter maximal step lengths of older non-fallers and fallers, but step length correlated with every variable tested. Thus, declines in maximum step length could indicate declines in hip and knee extensor kinetics and impaired performance on similar tasks like recovering from a trip.


Respiratory aspects of neurological disease.
Neurological disease may result in respiratory dysfunction; however the manifestations of respiratory dysfunction in such patients may be atypical because of wider effects of their underlying condition. In the present review we have considered separately acute neuromuscular respiratory disease (as well as aspects of respiratory muscle function relevant to intensive care), chronic neuromuscular respiratory disease, sleep related disorders, respiratory consequences of specific neurological diseases, and neurological features of respiratory disease. Approaches to specific clinical problems are discussed; in many instances this can be expedited by close cooperation with a respiratory physician. We suggest that management of respiratory dysfunction in neurological disease depends critically on three factors: firstly, knowledge of when respiratory dysfunction is likely to occur; secondly, maintaining a high index of clinical suspicion (specifically apparently vague symptoms should not be uncritically attributed to the underlying neurological condition); and, thirdly, the pursuing of appropriate investigations.


Beta-thalassemia and the advent of new interventions beyond transfusion and iron chelation.
Beta-thalassaemia, including sickle cell anaemia and thalassaemia E, is most common in developing countries in tropical and subtropic regions. Because carriers have migrated there owing to demographic migration, β-thalassaemia can now be detected in areas other than malaria-endemic areas. Every year, an estimated 300 000-500 000 infants, the vast majority of whom are from developing countries, are born with a severe haemoglobin anomaly. Currently, some basic techniques, which include iron chelation therapy, hydroxyurea, blood transfusion, splenectomy and haematopoietic stem cell transplantation, are being used to manage thalassaemia patients. Despite being the backbone of treatment, traditional techniques have several drawbacks and limitations. Ineffective erythropoiesis, correction of globin chain imbalance and adjustment of iron metabolism are some of the innovative treatment methods that have been developed in the care of thalassaemia patients in recent years. Moreover, regulating the expression of B-cell lymphoma/leukaemia 11A and sex-determining region Y-box through the enhanced expression of micro RNAs can also be considered putative targets for managing haemoglobinopathies. This review focuses on the biological basis of β-globin gene production, the pathophysiology of β-thalassaemia and the treatment options that have recently been introduced.


An accurate assay for HCV based on real-time fluorescence detection of isothermal RNA amplification.
Hepatitis C virus (HCV) is one of the common reasons of liver fibrosis and hepatocellular carcinoma (HCC). Early, rapid and accurate HCV RNA detection is important to prevent and control liver disease. A simultaneous amplification and testing (SAT) assay, which is based on isothermal amplification of RNA and real-time fluorescence detection, was designed to optimize routine HCV RNA detection. In this study, HCV RNA and an internal control (IC) were amplified and analyzed simultaneously by SAT assay and detection of fluorescence using routine real-time PCR equipment. The assay detected as few as 10 copies of HCV RNA transcripts. We tested 705 serum samples with SAT, among which 96.4% (680/705) showed consistent results compared with routine real-time PCR. About 92% (23/25) discordant samples were confirmed to be same results as SAT-HCV by using a second real-time PCR. The sensitivity and specificity of SAT-HCV assay were 99.6% (461/463) and 100% (242/242), respectively. In conclusion, the SAT assay is an accurate test with a high specificity and sensitivity which may increase the detection rate of HCV. It is therefore a promising tool to diagnose HCV infection.


Early Holocene human presence in Madagascar evidenced by exploitation of avian megafauna.
Previous research suggests that people first arrived on Madagascar by ~2500 years before present (years B.P.). This hypothesis is consistent with butchery marks on extinct lemur bones from ~2400 years B.P. and perhaps with archaeological evidence of human presence from ~4000 years B.P. We report >10,500-year-old human-modified bones for the extinct elephant birds Aepyornis and Mullerornis, which show perimortem chop marks, cut marks, and depression fractures consistent with immobilization and dismemberment. Our evidence for anthropogenic perimortem modification of directly dated bones represents the earliest indication of humans in Madagascar, predating all other archaeological and genetic evidence by >6000 years and changing our understanding of the history of human colonization of Madagascar. This revision of Madagascar's prehistory suggests prolonged human-faunal coexistence with limited biodiversity loss.


Prevalence of small for gestational age (SGA) and short stature in children born SGA who qualify for growth hormone treatment at 3 years of age: Population-based study.
To treat children born small for gestational age (SGA) with severe short stature, treatment with growth hormone (GH) has been approved in the USA, Europe, and Japan, but no population-based studies have reported their prevalence. The aims of this study were to investigate the prevalence of SGA and short stature in children born SGA who qualify for GH treatment at 3 years of age in a Japanese population. A population-based study was conducted in Kobe, Japan with 27 228 infants who were born between 2006 and 2008 and followed until 3 years of age. Prevalence of birthweight (BW) or birth length (BL) ≤ -2.0 standard deviation scores (SDS) for gestational age (GA; definition of SGA) was calculated. Short children born SGA who qualify for GH treatment at 3 years of age were estimated using the following criteria: BW and BL below the 10th percentile for GA, BW or BL ≤ -2.0 SDS for GA, and 2.5 SDS below the mean height for age. The prevalence of SGA was 3.5%. The estimated prevalence of short stature in children born SGA who met the criteria for GH treatment was 0.06%. The prevalence in infants born <34 weeks (0.39%) was significantly higher than that in infants born 34-41 weeks GA (0.05%, P = 0.02). The prevalence of SGA and short stature in children born SGA who qualify for GH treatment is approximately 1 of 30 infants and 1 of 1800 children, respectively. The risk is increased when children are born <34 weeks GA.


Lupus pathobiology based on genomics.
Systemic lupus erythematosus (SLE) is an autoimmune disorder with complex genetic underpinnings. This review attempts to assemble the myriad of genomic findings to build a clearer picture of the pathobiology of SLE to serve as a guide for therapeutics. Over 100 genes are now known for SLE, and several more penetrant ones have led to the emergence of more defined lupus phenotypes. Also discussed here are the targeted therapies that have come up on the horizon and the specific biologic mechanisms of more traditional therapies which have only recently been explored. The diagnostic toolbox has been enhanced by the addition of new antibodies, gene expression signatures, and mutation panels. This provides an opportunity to piece together the lupus puzzle and even revisit the clinical classification of SLE.


Development of SARS-CoV-2 packaged RNA reference material for nucleic acid testing.
Nucleic acid tests to detect the SARS-CoV-2 virus have been performed worldwide since the beginning of the COVID-19 pandemic. For the quality assessment of testing laboratories and the performance evaluation of molecular diagnosis products, reference materials (RMs) are required. In this work, we report the production of a lentiviral SARS-CoV-2 RM containing approximately 12 kilobases of its genome including common diagnostics targets such as RdRp, N, E, and S genes. The RM was measured with multiple assays using two different digital PCR platforms. To measure the homogeneity and stability of the lentiviral SARS-CoV-2 RM, reverse transcription droplet digital PCR (RT-ddPCR) was used with in-house duplex assays. The copy number concentration of each target gene in the extracted RNA solution was then converted to that of the RM solution. Their copy number values are measured to be from 1.5 × 105 to 2.0 × 105 copies/mL. The RM has a between-bottle homogeneity of 4.80-8.23% and is stable at 4 °C for 1 week and at -70 °C for 6 months. The lentiviral SARS-CoV-2 RM closely mimics real samples that undergo identical pre-analytical processes for SARS-CoV-2 molecular testing. By offering accurate reference values for the absolute copy number of viral target genes, the developed RM can be used to improve the reliability of SARS-CoV-2 molecular testing.