Stress and its timing: critical factors in determining the consequences of dopaminergic agents.
Dopaminergic agonists and antagonists can have opposite effects, depending on the state of the organism involved. The importance of prior state in determining the consequences of treatment with dopaminergic agents is illustrated in Serra's recent experiments. Although repeated treatment with DA antagonists enhanced the hypokinetic effects of 10 micrograms/kg of apomorphine, they obviated the influence of 25 micrograms/kg. In the former case, chronic DA-receptor blockade induced supersensitivity of DA autoreceptors; in the latter it resulted in subsensitivity. Since autoreceptor subsensitivity typically occurs after repeated treatment with DA agonists these experiments illustrate that neuroleptics caN behave like DA agonists. We discuss these and similar studies, emphasizing the role of stress and time in determining the consequences of dopaminergic agents.


Community gene annotation in practice.
Manual annotation of genomic data is extremely valuable to produce an accurate reference gene set but is expensive compared with automatic methods and so has been limited to model organisms. Annotation tools that have been developed at the Wellcome Trust Sanger Institute (WTSI, http://www.sanger.ac.uk/.) are being used to fill that gap, as they can be used remotely and so open up viable community annotation collaborations. We introduce the 'Blessed' annotator and 'Gatekeeper' approach to Community Annotation using the Otterlace/ZMap genome annotation tool. We also describe the strategies adopted for annotation consistency, quality control and viewing of the annotation. DATABASE URL: http://vega.sanger.ac.uk/index.html.


Comparison of clinical outcomes between endoscopic and radiologic placement of self-expandable metal stent in patients with malignant colorectal obstruction.
This study compared the clinical outcomes between endoscopic and radiologic placement of self-expandable metal stent (SEMS) in patients with malignant colorectal obstruction. In total, 111 patients were retrospectively enrolled in this study between January 2003 and June 2011 at Seoul National University Boramae Hospital. Technical and clinical success rates, complication rates, and stent patency were compared between using an endoscopic (n=73) or radiologic (n=38) method during the SEMS placement procedure. The technical success rate was higher in the endoscopic method than in the radiologic method (100% [73/73] vs. 92.1% [35/38], respectively; p=0.038). In addition, in 3 of the remaining 35 patients in the radiologic-method group, adjuvant endoscopic assistance was required. In the six patients (including the three aforementioned patients), the causes of technical failure were the inability to pass the guidewire into an obstructive lesion due to a tortuous, curved angulation of the sigmoid or descending colon (n=4), and a difficult approach to a lesion located at the descending or transverse colon (n=2). The clinical success rate, complication rate, and stent patency did not differ significantly between the two methods (p=0.424, 0.303, and 0.423, respectively). When the colorectal obstruction had a tortuous, curved angulation of the colon or was located at or proximal to the descending colon, the endoscopic method of SEMS placement appears to be more useful than the radiologic method. However, once SEMS placement was technically successful, the clinical success rate, complication rate, and stent patency did not differ with the method of insertion.


Role of Chromodomain-Helicase-DNA-Binding Protein 4 (CHD4) in Breast Cancer.
Chromodomain-helicase-DNA-binding protein 4 (CHD4) is an epigenetic regulator identified as an oncogenic element that may provide a novel therapeutic target for the treatment of breast cancer (BC). CHD4-the core component of the nucleosome remodeling and deacetylase (NuRD) complex-may be mutated in patients with this disease. However, information on CHD4 mutants that might allow their use as biomarkers of therapeutic success and prognosis is lacking. The present work examines mutations in CHD4 reported in patients with breast cancer and included in public databases and attempts to identify their roles in its development. The databases revealed 81 point mutations across different types of breast cancer (19 of which also appeared in endometrial, intestinal, nervous system, kidney, and lymphoid organ cancers). 71.6% of the detected mutations were missense mutations, 13.6% were silent, and 6.2% nonsense. Over 50% affected conserved residues of the ATPase motor (ATPase and helicase domains), and domains of unknown function in the C-terminal region. Thirty one mutations were classified in the databases as either 'deleterious', 'probably/possibly damaging' or as 'high/medium pathogenic'; another five nonsense and one splice-site variant were predicted to produce potentially harmful truncated proteins. Eight of the 81 mutations were categorized as putative driver mutations and have been found in other cancer types. Some mutations seem to influence ATPase and DNA translocation activities (R1162W), while others may alter protein stability (R877Q/H, R975H) or disrupt DNA binding and protein activity (R572*, X34_splice) suggesting CHD4 function may be affected. In vivo tumorigenecity studies in endometrial cancer have revealed R975H and R1162W as mutations that lead to CHD4 loss-of-function. Our study provides insight into the molecular mechanism whereby CHD4, and some of its mutants could play a role in breast cancer and suggest important implications for the biological comprehension and prognosis of breast cancer, identifying CHD4 as a novel therapeutic target for BC patients.


Schistocytic hemolytic anemia owing to central venous catheter in a child with acute lymphoblastic leukemia.
A 4-year-old boy with acute lymphoblastic leukemia (ALL) was inserted a central venous catheter into right vena jugularis interna, whereas on BFM-ALL (Protocol 1) therapy. He developed progressive anemia, thrombocytopenia, schistocytes with triangular or crescent shapes, microspherocytes and teardrop cells in the peripheric blood smear, elevated lactate dehydrogenase, and decreased haptoglobin. Schistocytic hemolytic anemia was considered. On chest radiogram, the central venous catheter (CVC) tip was seen beneath the expected location. An echocardiography revealed that CVC was within the right atrium, in contact with tricuspite leaflets. So, CVC was overhauled and a new one was inserted. After the revision, hemolysis ceased; haptoglobin level normalized.


One-day workflow scheme for bacterial pathogen detection and antimicrobial resistance testing from blood cultures.
Bloodstream infections are associated with high mortality rates because of the probable manifestation of sepsis, severe sepsis and septic shock(1). Therefore, rapid administration of adequate antibiotic therapy is of foremost importance in the treatment of bloodstream infections. The critical element in this process is timing, heavily dependent on the results of bacterial identification and antibiotic susceptibility testing. Both of these parameters are routinely obtained by culture-based testing, which is time-consuming and takes on average 24-48 hours(2, 4). The aim of the study was to develop DNA-based assays for rapid identification of bloodstream infections, as well as rapid antimicrobial susceptibility testing. The first assay is a eubacterial 16S rDNA-based real-time PCR assay complemented with species- or genus-specific probes(5). Using these probes, Gram-negative bacteria including Pseudomonas spp., Pseudomonas aeruginosa and Escherichia coli as well as Gram-positive bacteria including Staphylococcus spp., Staphylococcus aureus, Enterococcus spp., Streptococcus spp., and Streptococcus pneumoniae could be distinguished. Using this multiprobe assay, a first identification of the causative micro-organism was given after 2 h. Secondly, we developed a semi-molecular assay for antibiotic susceptibility testing of S. aureus, Enterococcus spp. and (facultative) aerobe Gram-negative rods(6). This assay was based on a study in which PCR was used to measure the growth of bacteria(7). Bacteria harvested directly from blood cultures are incubated for 6 h with a selection of antibiotics, and following a Sybr Green-based real-time PCR assay determines inhibition of growth. The combination of these two methods could direct the choice of a suitable antibiotic therapy on the same day (Figure 1). In conclusion, molecular analysis of both identification and antibiotic susceptibility offers a faster alternative for pathogen detection and could improve the diagnosis of bloodstream infections.


Evaluation of Medication Instruction Sheets for Patients Undergoing R-CHOP Therapy in Non-Hodgkin's Lymphoma.
High-dose chemotherapy is frequently administered to patients with hematologic malignancies, thereby causing severe adverse drug reactions (ADRs) at a relatively high frequency. To precisely monitor ADRs, we developed a medication instruction sheet (MIS) for patients who received rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone (R-CHOP) combination therapy for non-Hodgkin's lymphoma (NHL). Herein, we evaluated the usefulness of the MIS for managing ADRs in patients who received R-CHOP therapy. We included patients aged ≥20 years who received R-CHOP therapy as first-line treatment for NHL at the Department of Hematology, Kyushu University Hospital, between August 2014 and December 2018. Medical professionals evaluated the possible occurrence of ADRs according to the present MIS and ADRs were graded according to the Common Toxicity Criteria, version 4.0 (National Cancer Institute, Bethesda, MD, USA). Finally, the accuracy of the MIS in predicting the occurrence of ADRs of different grades and during definite periods was evaluated. Seventy-five patients with NHL were included in the present study. Overall, 359 ADR events were monitored, which were predicted ADR items listed in the MIS. Among these, 254 (71%) events occurred during the same period as those listed in the MIS. The onset timing of any grade of an infusion reaction and peripheral neuropathy precisely matched those listed in the MIS. However, the accuracy of the MIS was reduced in patients with thrombocytopenia (42%). The present MIS could be useful for monitoring ADRs in patients with cancer undergoing R-CHOP therapy.


Effect of nucleobase change on cytosine deamination through DNA photo-cross-linking reaction via 3-cyanovinylcarbazole nucleoside.
Photo-chemical deamination of cytosine using 3-cyanovinylcarbazole nucleoside (CNVK) mediated photo-cross-linking is a technique for site-directed mutagenesis. Using this technique in vivo requires the elimination of a high-temperature incubation step; instead, incubation should be carried out under physiological conditions. To improve the reactivity of CNVK mediated photo-cross-link induced deamination of cytosine under physiological conditions, an evaluation of base pairing in cytosine was carried out with respect to its deamination. Guanine was replaced with 4 different counter bases (inosine, 2-aminopurine, 5-nitroindole, and nebularine), showing distinct hydrogen bonding patterns with target cytosine, which was incorporated at the -1 position with respect to CNVK in the CNVK-modified photo-responsive oligodeoxyribonucleotides to ascertain the role of hydrogen bonding in deamination under physiological conditions. Among the counter bases, inosine showed the highest acceleration towards the photo-induced deamination reaction.


MFN1 structures reveal nucleotide-triggered dimerization critical for mitochondrial fusion.
Mitochondria are double-membraned organelles with variable shapes influenced by metabolic conditions, developmental stage, and environmental stimuli. Their dynamic morphology is a result of regulated and balanced fusion and fission processes. Fusion is crucial for the health and physiological functions of mitochondria, including complementation of damaged mitochondrial DNAs and the maintenance of membrane potential. Mitofusins are dynamin-related GTPases that are essential for mitochondrial fusion. They are embedded in the mitochondrial outer membrane and thought to fuse adjacent mitochondria via combined oligomerization and GTP hydrolysis. However, the molecular mechanisms of this process remain unknown. Here we present crystal structures of engineered human MFN1 containing the GTPase domain and a helical domain during different stages of GTP hydrolysis. The helical domain is composed of elements from widely dispersed sequence regions of MFN1 and resembles the 'neck' of the bacterial dynamin-like protein. The structures reveal unique features of its catalytic machinery and explain how GTP binding induces conformational changes to promote GTPase domain dimerization in the transition state. Disruption of GTPase domain dimerization abolishes the fusogenic activity of MFN1. Moreover, a conserved aspartate residue trigger was found to affect mitochondrial elongation in MFN1, probably through a GTP-loading-dependent domain rearrangement. Thus, we propose a mechanistic model for MFN1-mediated mitochondrial tethering, and our results shed light on the molecular basis of mitochondrial fusion and mitofusin-related human neuromuscular disorders.


Transcription factor AP4 modulates reversible and epigenetic silencing of the Cd4 gene.
CD4 coreceptor expression is negatively regulated through activity of the Cd4 silencer in CD4(-)CD8(-) double-negative (DN) thymocytes and CD8(+) cytotoxic lineage T cells. Whereas Cd4 silencing is reversed during transition from DN to CD4(+)CD8(+) double-positive stages, it is maintained through heritable epigenetic processes following its establishment in mature CD8(+) T cells. We previously demonstrated that the Runx family of transcription factors is required for Cd4 silencing both in DN thymocytes and CD8(+) T cells. However, additional factors that cooperate with Runx proteins in the process of Cd4 silencing remain unknown. To identify collaborating factors, we used microarray and RNAi-based approaches and found the basic helix-loop-helix ZIP transcription factor AP4 to have an important role in Cd4 regulation. AP4 interacts with Runx1 in cells in which Cd4 is silenced, and is required for Cd4 silencing in immature DN thymocytes through binding to the proximal enhancer. Furthermore, although AP4-deficient CD8(+) T cells appeared to normally down-regulate CD4 expression, AP4 deficiency significantly increased the frequency of CD4-expressing effector/memory CD8(+) T cells in mice harboring point mutations in the Cd4 silencer. Our results suggest that AP4 contributes to Cd4 silencing both in DN and CD8(+) T cells by enforcing checkpoints for appropriate timing of CD4 expression and its epigenetic silencing.


Effect of distance between homologous sequences and 3' homology on the frequency of retroviral reverse transcriptase template switching.
Deletion of direct repeats in retroviral genomes provides an in vivo system for analysis of reverse transcriptase (RT) template switching. The effect of distance between direct repeats on the rate of deletion was determined for 16 murine leukemia virus (MLV)-based vectors containing a 701-bp direct repeat of overlapping fragments of the herpes simplex virus thymidine kinase gene (HTK). The direct repeats were separated by spacer fragments of various lengths (0.1 to 3.5 kb). Southern analysis of infected cells after one replication cycle indicated that all vectors in which the distance between homologous sequences was >1,500 bp deleted at very high rates (>90%). In contrast, vectors containing <1,500 bp between homologous sequences exhibited lower frequencies of deletion (37 to 82%). To analyze the pattern of locations at which RT switched templates, restriction site markers were introduced to divide the downstream direct repeat into five regions. RT switched templates within all five regions of the 701-bp direct repeat and the frequency of template switching was greater within the 5' regions in comparison to the 3' regions. The probability of RT switching templates within the 5' regions doubled when the MLV packaging sequence (Psi) was placed between the 701-bp direct repeats. However, Psi did not increase the rate of template switching for shorter direct repeats. These results indicate that linear distance between homologous sequences increases the rate of template switching and suggest that duplex formation between nascent DNA and homologous template sequences 3' of RT promote template switching.


Effect of rhuIFN-gamma treatment in multibacillary leprosy patients.
Previous studies have shown that when multibacillary leprosy patients were treated with recombinant human interferon gamma (rhuIFN-gamma) for 6-10 months there was an accelerated reduction in the number of acid-fast bacilli in the skin at the site of injection as well as an accelerated bacillary reduction at distal sites. However, this favorable out-come of IFN-gamma treatment was associated with the development of erythema nodosum leprosum (ENL). The present study was undertaken to investigate whether rhuIFN-gamma-induced bacillary clearance could be disassociated from the induction of ENL. rhuIFN-gamma was administered together with thalidomide and conventional multidrug chemotherapy to newly diagnosed leprosy patients. During treatment with this combination of drugs, the mean reduction in bacterial load was the same as the reduction observed with chemotherapy alone. Moreover, the inclusion of thalidomide in the treatment regimen was associated with a low frequency of ENL episodes. A second group of leprosy patients, who had already completed 2 years of chemotherapy, were treated with rhuIFN-gamma only. In those patients who were skin bacilli negative, ENL did not occur during rhuIFN-gamma treatment. In contrast, in bacilli-positive patients the frequency of ENL during rhuIFN-gamma treatment was higher, as was the occurrence of local erythema and induration. However, rhuIFN-gamma treatment without concomitant chemotherapy did not result in a reduction in the bacterial load in the skin of bacilli-positive patients. These findings, taken together, indicate that rhuIFN-gamma does not, by itself, accelerate bacterial clearance, but requires concomitant chemotherapy to achieve the accelerated reduction in bacillary load. Thalidomide reduces the frequency of IFN-gamma-induced ENL, but also eliminates the IFN-gamma-induced bacillary clearance.


Home sampling for sexually transmitted infections and HIV in men who have sex with men: a prospective observational study.
To determine uptake of home sampling kit (HSK) for STI/HIV compared to clinic-based testing, whether the availability of HSK would increase STI testing rates amongst HIV infected MSM, and those attending a community-based HIV testing clinic compared to historical control. Prospective observational study in three facilities providing STI/HIV testing services in Brighton, UK was conducted. Adult MSM attending/contacting a GUM clinic requesting an STI screen (group 1), HIV infected MSM attending routine outpatient clinic (group 2), and MSM attending a community-based rapid HIV testing service (group 3) were eligible. Participants were required to have no symptomatology consistent with STI and known to be immune to hepatitis A and B (group 1). Eligible men were offered a HSK to obtain self-collected specimens as an alternative to routine testing. HSK uptake compared to conventional clinic-based STI/HIV testing in group 1, increase in STI testing rates due to availability of HSK compared to historical controls in group 2 and 3, and HSK return rates in all settings were calculated. Among the 128 eligible men in group 1, HSK acceptance was higher (62.5% (95% CI: 53.5-70.9)) compared to GUM clinic-based testing (37.5% (95% CI: 29.1-46.5)), (p = 0.0004). Two thirds of eligible MSM offered an HSK in all three groups accepted it, but HSK return rates varied (highest in group 1, 77.5%, lowest in group 3, 16%). HSK for HIV testing was acceptable to 81% of men in group 1. Compared to historical controls, availability of HSK increased the proportion of MSM testing for STIs in group 2 but not in group 3. HSK for STI/HIV offers an alternative to conventional clinic-based testing for MSM seeking STI screening. It significantly increases STI testing uptake in HIV infected MSM. HSK could be considered as an adjunct to clinic-based services to further improve STI/HIV testing in MSM.


Apolipoprotein E and alpha-1-antichymotrypsin allele polymorphism in sporadic and familial Alzheimer's disease.
Alzheimer disease (AD) patients with both sporadic and familial forms of AD and non-demented controls were genotyped for common polymorphisms in the signal peptide for alpha-1-antichymotrypsin (ACT) gene and in two different regions of apolipoprotein E (APOE) gene. The ACT TT genotype was over-represented (P = 0.025) in patients with early onset of sporadic AD. In this patient's group ACT TT genotype conferred a significant crude odds ratio for the disease (OR = 2.09; 95% CI = 1.09-4.00, P = 0.025). After adjustment for the APOE epsilon4 and APOE -491 genotypes, logistic regression analysis confirmed that the ACT TT genotype resulted independently associated with early onset AD (adjusted OR = 2.56; 85% CI = 1.3-5.2, P = 0.009). The frequency of APOE epsilon4 allele was increased in AD, as expected (OR = 5.92, 95% CI = 3.60-9.70, P = 0.0001). On the contrary, the APOE -491 A/T genotypes were not associated with AD. No preferential association of the APOE epsilon4 allele or APOE -491 A/T genotypes with ACT A/T alleles was observed in AD. Present findings indicated that subjects with ACT TT genotype had an increased risk of developing AD and suggested that this genotype influenced the risk of an early onset of the disease by affecting the production of ACT molecules.


Development and test of highly accurate endpoint free energy methods. 3: partition coefficient prediction using a Poisson-Boltzmann method combined with a solvent accessible surface area model for SAMPL challenges.
Accurately predicting solvation free energy is the key to predict protein-ligand binding free energy. In addition, the partition coefficient (log P), which is an important physicochemical property that determines the distribution of a drug in vivo, can be derived directly from transfer free energies, i.e., the difference between solvation free energies (SFEs) in different solvents. Within the Statistical Assessment of the Modeling of Proteins and Ligands (SAMPL) 9 challenge, we applied the Poisson-Boltzmann (PB) surface area (SA) approach to predict the toluene/water transfer free energy and partition coefficient (log Ptoluene/water) from SFEs. For each solute, only a single conformation automatically generated by the free software Open Babel was used. The PB calculation directly adopts our previously optimized boundary definition - a set of general AMBER force field 2 (GAFF2) atom-type based sphere radii for solute atoms. For the non-polar SA model, we newly developed the solvent-related molecular surface tension parameters γ and offset b for toluene and cyclohexane targeting experimental SFEs. This approach yielded the highest predictive accuracy in terms of root mean square error (RMSE) of 1.52 kcal mol-1 in transfer free energy for 16 small drug molecules among all 18 submissions in the SAMPL9 blind prediction challenge. The re-evaluation of the challenge set using multi-conformation strategies based on molecular dynamics (MD) simulations further reduces the prediction RMSE to 1.33 kcal mol-1. At the same time, an additional evaluation of our PBSA method on the SAMPL5 cyclohexane/water distribution coefficient (log Dcyclohexane/water) prediction revealed that our model outperformed COSMO-RS, the best submission model with RMSEPBSA = 1.88 versus RMSECOSMO-RS = 2.11 log units. Two external log Ptoluene/water and log Pcyclohexane/water datasets that contain 110 and 87 data points, respectively, are collected for extra validation and provide an in-depth insight into the error source of the PBSA method.


Effects of gamma-irradiation on the infectivity and chromosome aberration of Clonorchis sinensis.
Effects of gamma irradiation on the worm survival and chromosomal aberration of Clonorchis sinensis were studied. The metacercariae irradiated with various amounts of gamma radiation (ranging from 5 Gy to 50 Gy) were fed to rats, and the effects were compared with those of non-irradiated controls. Recovery rates of adult worms in irradiated groups were reduced gradually as increasing of the irradiation doses. No worm was recovered from rats which were fed with 50 Gy irradiated metacercariae. The chromosome number was 2n = 56 in all worms from all experimental groups. However, the groups irradiated with 20 Gy, 25 Gy or 30 Gy showed variations in the chromosome number, depending on different cells in the same individual. Radiation doses used in this study did not appear to induce chromosome aberrations, however, irradiation with 30 Gy showed slightly reduced chromosome size.


Butanol Purified Food Allergy Herbal Formula-2 Has an Immunomodulating Effect ex-vivo in Pediatric Crohn's Disease Subjects.
Background: TNF-α has a major role in the pathogenesis of Crohn's disease (CD). In contrast, GM-CSF may be beneficial for its anti-inflammatory role in a subset of patients with CD with antibodies against GM-CSF as seen in prior trials of GM-CSF which resulted in clinical improvement in CD. We developed butanol purified Food Allergy Herbal Formula-2 (B-FAHF-2) by refining FAHF-2. FAHF-2 suppressed TNF-α production by human peripheral blood mononuclear cells (PBMCs) and colonic mucosa, and abrogated colitis in a murine model. We sought to examine the effect of B-FAHF-2 and the herbs that comprise it on TNF-α and GM-CSF production as a potential herbal therapy for the treatment of CD. Methods: B-FAHF-2 was examined using high pressure liquid chromatography (HPLC) and compared to the original formulation, FAHF-2. PBMCs from pediatric patients with CD were cultured with lipopolysaccharide and B-FAHF-2, individual herbs or medium alone. Colonic biopsy specimens were cultured with or without B-FAHF-2. TNF-α and GM-CSF were measured by enzyme-linked immunosorbent assay (ELISA). B-FAHF-2 efficacy was tested in vivo in the CD45Rbhi transfer model. Results: B-FAHF-2 had a similar HPLC fingerprint as FAHF-2 but decreased TNF-α production by PBMCs and colonic mucosa from pediatric CD subjects at 20% of the FAHF-2 dose. B-FAHF-2 increased GM-CSF production by PBMCs and colonic mucosa from pediatric CD subjects including those with antibodies to GM-CSF. Of B-FAHF-2's herbal constituents, only Huang Bai suppressed TNF-α and increased GM-CSF production. In the murine model, B-FAHF-2 treatment alleviated colitis. Conclusions: B-FAHF-2 decreased TNF-α production by PBMCs and colonic mucosa from pediatric subjects at a lower dose than FAHF-2. B-FAHF-2 also increased GM-CSF production by PBMCs independent of antibodies. B-FAHF-2 may have a benefit in CD patients.


Role of A2a extracellular adenosine receptor-mediated signaling in adenosine-mediated inhibition of T-cell activation and expansion.
Accumulation of adenosine and of deoxyadenosine in the absence of adenosine deaminase activity (ADA) activity results in lymphocyte depletion and in severe combined immunodeficiency (ADA SCID), which is currently explained by direct cell death-causing effects of intracellular products of adenosine metabolism. We explored the alternative mechanisms of peripheral T-cell depletion as due to inhibition of T-cell expansion by extracellular adenosine-mediated signaling through purinergic receptors. The strong inhibition of the T-cell receptor (TCR)-triggered proliferation and of upregulation of interleukin-2 receptor alpha chain (CD25) molecules, but not the direct lymphotoxicity, were observed at low concentrations of extracellular adenosine. These effects of extracellular adenosine (Ado) are likely to be mediated by A2a receptor-mediated signaling rather than by intracellular toxicity of adenosine catabolites, because (1) poorly metabolized adenosine analogs cause the accumulation of cAMP and strong inhibition of TCR-triggered CD25 upregulation; (2) the A2a, but not the A1 or A3, receptors are the major expressed and functionally coupled adenosine receptors in mouse peripheral T and B lymphocytes, and the adenosine-induced cAMP accumulation in lymphocytes correlates with the expression of A2a receptors; (3) the specific agonist of A2a receptor, CGS21680, induces increases in [cAMP]i in lymphocytes, whereas the specific antagonist of A2a receptor, CSC, inhibits the effects of Ado and CGS21680; and (4) the increases in [cAMP]i mimic the adenosine-induced inhibition of TCR-triggered CD25 upregulation and splenocyte proliferation. These studies suggest the possible role of adenosine receptors in the regulation of lymphocyte expansion and point to the downregulation of A2a purinergic receptors on T cells as a potentially attractive pharmacologic target.


Reduction of bias in the evaluation of fractional anisotropy and mean diffusivity in magnetic resonance diffusion tensor imaging using region-of-interest methodology.
Accurate quantification of fractional anisotropy (FA) and mean diffusivity (MD) in MR diffusion tensor imaging (DTI) requires adequate signal-to-noise ratio (SNR) especially in low FA areas of the brain, which necessitates clinically impractical long image acquisition times. We explored a SNR enhancement strategy using region-of-interest (ROI)-based diffusion tensor for quantification. DTI scans from a healthy male were acquired 15 times and combined into sets with different number of signal averages (NSA = 1-4, 15) at one 1.5-T Philips and three 3-T (Philips, Siemens and GE) scanners. Equivalence test was performed to determine NSA thresholds for bias-free FA and MD quantifications by comparison with reference values derived from images with NSA = 15. We examined brain areas with low FA values including caudate nucleus, globus pallidus, putamen, superior temporal gyrus, and substructures within thalamus (lateral dorsal, ventral anterior and posterior nuclei), where bias-free FA is difficult to obtain using a conventional approach. Our results showed that bias-free FA can be obtained with NSA = 2 or 3 in some cases using ROI-based analysis. ROI-based analysis allows reliable FA and MD quantifications in various brain structures previously difficult to study with clinically feasible data acquisition schemes.


Antibody glycosylation in pregnancy and in newborns: biological roles and implications.
Glycosylation patterns have the potential to affect the function of antibody, antibody half-life and transplacental transfer from mother to foetus. Here, we review recent advances in our understanding of how glycosylation patterns of antibodies may be altered during pregnancy, vaccination and infection. During pregnancy, there is preferential transplacental transfer of natural killer (NK) cell-activating antibodies that are galactosylated and sialylated, against both bacterial and viral antigens. Markers of NK cell function are also associated with a higher abundance of galactosylation and sialylation in respiratory syncytial virus-specific IgG, compared with total IgG, in infants up to 7 months of age which may suggest a role for NK-cell activating antibodies as important mediators of immunity during early infancy. Differential glycosylation patterns have been observed in some respiratory conditions, as increased nongalactosylated antibodies have been associated with the development of chronic inflammatory bronchopulmonary dysplasia (BPD) in preterm infants. Glycosylation patterns in children appear age-dependent, which could modulate the effector function of IgG. The clinical relevance of these findings needs to be established. Glycosylation plays a key role in mediating antibody function. Glycosylation patterns associated with positive outcomes from infection in mothers and infants could inform the design of the next generation of vaccines for use in pregnancy and infancy. SDC VIDEO LINK:: http://links.lww.com/COID/A29.


Is the hemidesmosome a half desmosome? An immunological comparison of mammalian desmosomes and hemidesmosomes.
Although the mammalian epidermal basal cell hemidesmosome bears some superficial resemblance to one half of a desmosome at the ultrastructural level, examination of the structure of the electron-dense submembranous plaques of the hemidesmosome and desmosome reveals that they differ with respect to their overall morphology and dimensions. Based on these findings, we wondered whether components of the desmosome are present in the hemidesmosome. In order to determine this we prepared a number of stratified squamous epithelial tissues for indirect immunofluorescence using antibody preparations directed against known desmosome components including desmoplakin and certain glycoproteins. These antibody preparations do not show reaction with hemidesmosomes by indirect immunofluorescence criteria. We have also utilized bullous pemphigoid (BP) autoantibodies that have been shown to recognize hemidesmosomes in mammalian skin cells [Mutasim et al., J. Invest. Derm., 84:47-53, 1985]. Double label indirect immunofluorescence observations of neonatal mouse skin prepared using desmoplakin antibodies and BP autoantibodies reveal that hemidesmosomes that are stained by the BP autoantibodies are not recognized by the desmoplakin antibodies. We confirmed these findings at the ultrastructural level by indirect immunogold localization of desmoplakin antibodies and BP autoantibodies. Therefore, the hemidesmosome does not appear to be one half of a desmosome and may possess a very different molecular organization relative to the desmosome. We raise the possibility that the variability between the hemidesmosome and desmosome that we detect at the morphological and immunological level may reflect the functional differences of these two types of junctions.


Ki-67/MKI67 as a Predictive Biomarker for Clinical Outcome in Gastric Cancer Patients: an Updated Meta-analysis and Systematic Review involving 53 Studies and 7078 Patients.
Gastric cancer (GC) threatens human health worldwide and we performed this meta-analysis to evaluate the clinical value of Ki-67/MKI67 in patients with GC. The combined hazard ratio (HR), odds ratio (OR) and 95% confidence interval (95% CI) were calculated to assess the relationships of Ki-67/MKI67 expression with prognoses and clinicopathological characteristics. Genes co-expressed with MKI67 were collected for Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and protein-protein interaction (PPI) network analyses. In total, 53 studies with 7078 patients were included in this study. The pooled HRs indicated that an elevated expression of Ki-67/MKI67 predicted an unfavorable overall survival (HR: 1.54, 95% CI: 1.33-1.78, P<0.0001) and disease-free survival (HR: 2.28, 95% CI: 1.43-3.64, P<0.0001) in GC patients. Additionally, in patients with advanced GC, a high Ki-67/MKI67 expression was also significantly connected with OS (HR: 1.37, 95% CI: 1.18-1.60, P<0.0001). The combined ORs showed that Ki-67/MKI67 expression was related to TNM stage (stage III/IV versus stage I/II: OR=1.93, 95% CI=1.34-2.78, P<0.0001), tumor differentiation (poor versus well/moderate: OR=1.94, 95% CI=1.32-2.85, P=0.001), lymph node metastasis (yes versus no: OR=1.67, 95% CI=1.23-2.25, P=0.001), distant metastasis (yes versus no: OR=1.67, 95% CI=1.24-2.26, P=0.001) and tumor invasion depth (T3/T4 versus Tis/T1/T2: OR=1.98, 95% CI=1.60-2.44, P<0.0001). The results of GO, KEGG pathway and PPI network analyses indicated that Ki-67/MKI67 may be involved in the development of GC via influencing P53 signaling pathway. Ki-67/MKI67 could be a potential indicator to predict the prognosis of patients with GC and identify high-risk cases. Detecting Ki-67/MKI67 expression in clinic may be helpful in optimizing individual treatment and further improving the survival expectancy of patients with GC.


Skin assessment of patients at risk of pressure ulcers.
The incidence of pressure ulcers in the community and in acute settings is a concern for patients and healthcare professionals. The high cost to healthcare services of treating individuals with pressure ulcers means prevention is much cheaper than cure. Nurses have a responsibility to implement and participate in prevention programmes to decrease the incidence of pressure ulcers. Assessment is the first stage in prevention. This article focuses on the assessment of pressure areas, providing the reader with the knowledge of where and when skin assessment should take place and what needs to be assessed.


Bronchiolar Adenoma: Expansion of the Concept of Ciliated Muconodular Papillary Tumors With Proposal for Revised Terminology Based on Morphologic, Immunophenotypic, and Genomic Analysis of 25 Cases.
We have identified 25 lesions involving alveolar lung parenchyma characterized by nodular proliferation of bland bilayered bronchiolar-type epithelium containing a continuous layer of basal cells. These lesions shared some histologic features with the recently described entity of ciliated muconodular papillary tumor (CMPT); however, the majority did not fit all diagnostic criteria in that they exhibited only focal or absent papillary architecture, and they had variable number of ciliated and mucinous cells, with some lesions entirely lacking 1 or both of these components. The morphologic and immunohistochemical features ranged from those resembling proximal bronchioles (proximal-type: moderate to abundant mucinous and ciliated cells; negative or weak TTF1 in luminal cells; n=8) to those resembling respiratory bronchioles (distal-type: scant or absent mucinous and ciliated cells; positive TTF1 in luminal cells; n=17). The hallmark of all lesions was a continuous layer of basal cells (p40 and CK5/6-positive). We provisionally designated these lesions as bronchiolar adenomas (BAs) and analyzed their clinicopathologic and molecular features. All BAs were discrete, sharply circumscribed lesions with a median size of 0.5 cm (range, 0.2 to 2.0 cm). Most lesions were either entirely flat (n=14) or contained focal papillary architecture (n=7); only 4 lesions, all proximal-type, were predominantly papillary, fitting the classic description of CMPT. Notably, of 9 lesions submitted for frozen section evaluation, 7 were diagnosed as adenocarcinoma. No postsurgical recurrences were observed for any lesions (median follow-up, 11 mo). Twenty-one BAs underwent next-generation sequencing and/or immunohistochemistry for BRAF V600E, revealing mutation profiles similar to those previously described for CMPTs, including BRAF V600E mutations (n=8, 38%), unusual EGFR exon 19 deletions (n=2, 10%), EGFR exon 20 insertions (n=2, 10%), KRAS mutations (n=5, 24%), and HRAS mutations (n=1, 5%). The mutation profiles were similar in proximal-type and distal-type lesions. In conclusion, we describe a family of putatively benign clonal proliferations with a spectrum of morphology recapitulating various levels of the bronchiolar tree, of which only a minor subset fits the classic description of CMPT. Comparable mutation profiles and partially overlapping morphologic features across the spectrum of these lesions support their nosological relationship. We propose designating this entire family of lesions as BAs, and that lesions currently designated CMPTs represent a subgroup of this family.


On the robustness of latent class models for diagnostic testing with no gold standard.
It is difficult to estimate sensitivity and specificity of diagnostic tests when there is no gold standard. Latent class models have been proposed as a potential solution as they provide estimates without the need for a gold standard. Using a motivating example of the evaluation of point of care tests for leptospirosis in Tanzania, we show how a realistic violation of assumptions underpinning the latent class model can lead directly to substantial bias in the estimates of the parameters of interest. In particular, we consider the robustness of estimates of sensitivity, specificity, and prevalence, to the presence of additional latent states when fitting a two-state latent class model. The violation is minor in the sense that it cannot be routinely detected with goodness-of-fit procedures, but is major with regard to the resulting bias.


Stereotactic pallidotomy for the treatment of Parkinson's disease. Efficacy and adverse effects at 6 months in 26 patients.
We evaluated the safety and efficacy of microelectrode-guided stereotactic pallidotomy in patients with advanced Parkinson's disease (PD). Using diagnostic criteria and evaluations outlined in the Core Assessment Programme in Transplantation (CAPIT) protocol, we studied unilateral pallidotomy in 26 patients with advanced idiophatic PD, motor fluctuations, and peak dose dyskinesias. All underwent unilateral stereotactic pallidotomy. Assessments conducted in the "practically defined off" and "best on" states at baseline and at 1 and 6 months postoperatively included Unified Parkinson's Disease Rating Scale (UPDRS) parts II, III, and IV and timed motor testing as outlined in CAPIT. Motor UPDRS in the "off" state improved at 1 and 6 months after surgery (p = 0.002, p = 0.008) Likewise, the sum of individual "off" contralateral motor UPDRS items improved (p = 0.0002, p = 0.0005). The duration (p = 0.0001 at 1 and p = 0.001 at 6 months) and severity (p = 0.003 at 1 and p = 0.0005 at 6 months) of dyskinesia improved, but other aspects of the "on" function were unchanged. Serious adverse effects occurred in eight patients and included one fatal deep and three nonfatal frontal lobe hemorrhages with resultant language or behavioral deficits. Nonhemorrhagic complications included one hemiparesis and three frontal lobe syndromes. Pallidotomy improves PD motor disability in the "off" state. Peak dose dyskinesias are reduced, although other aspects of "on" motor function are unchanged. Although morbidity may limit its use, pallidotomy is effective in targeting particular symptoms such as unremitting dyskinesia and severe "off" motor disability in advanced PD.


Cluster assembly route to a novel octagonal two-dimensional ZnO monolayer.
To explore the possibility of cluster assembly resulting in a two-dimensional (2D) stable monolayer of ZnO, a systematic study is performed on the structural evolution of bare and passivated stoichiometric clusters of [Formula: see text] [Formula: see text], [Formula: see text], using density-functional-theory-based first principles electronic structure calculations. The changes in hybridization are investigated with the aid of the site-projected partial density of states and partial charge density, while the effect of passivation and size on the ionicity of the cluster is studied using Bader charge analysis. The structural and chemical properties are found to be influenced by the coordination number of atoms in the clusters irrespective of the coordinating species. The physical parameters and hybridization of the states for the clusters on passivation resemble those of the bulk. Passivation thus provides an environment that leads to the stability of the clusters. Cluster assembly using the stable cluster geometries of passivated clusters (without the passivating atoms) has been shown to lead to stable 2D structures. This stability has been studied on the basis of binding energy, vibrational frequency, phonon dispersion and thermal properties. A new octagonal 2D monolayer planar geometry of ZnO is predicted.


Multiple forms of P700-chlorophyll a-protein complexes from Synechococcus sp.: The iron, quinone and carotenoid contents.
The iron, quinone and carotenoid contents of five P700-chlorophyll a-protein complexes having different subunit structures (CP1-a,-b,-c,-d and-e) from the thermophilic cyanobacterium Synechococcus sp. were determined. CP1-a,-b,-c and-d that commonly have four polypeptides of 62,000, 60,000, 14,000 and 10,000 dalton contained 10-14 iron atoms per P700, whereas CP1-e that lacks the two small polypeptides was totally devoid of iron. All CP1 complexes contained vitamin K1 at the molar ratio of vitamin K1 to P700 of about 2 except CP1-e that had only 0.4 vitamin K1 per P700. No plastoquinone was detected in five CP1 complexes. Out of four major carotenoids, β-carotene, zeaxanthin, caloxanthin, and myxoxanthophyll, present in the thylakoid membranes, only β-carotene was found in isolated CP1 complexes; all CP1 complexes contained about 10 β-carotene molecules per P700. The flourescence excitation spectrum showed that β-carotene serves as an efficient antenna of photosystem I. It is concluded that all iron atoms and a larger fraction of vitamin K1 molecules present in the photosystem I reaction center complex are associated with the 14,000 and 10,000 dalton polypeptides, whereas β-carotene exclusively binds to the large polypeptides which carry the functional and antenna chlorophyll a. The possible functions of iron and vitamin K1 as electron carriers and of β-carotene as the accessary pigment and a photoprotectant in the photosystem I complexes are discussed.


Regression supports two mechanisms of fork processing in phage T4.
Replication forks routinely encounter damaged DNA and tightly bound proteins, leading to fork stalling and inactivation. To complete DNA synthesis, it is necessary to remove fork-blocking lesions and reactivate stalled fork structures, which can occur by multiple mechanisms. To study the mechanisms of stalled fork reactivation, we used a model fork intermediate, the origin fork, which is formed during replication from the bacteriophage T4 origin, ori(34). The origin fork accumulates within the T4 chromosome in a site-specific manner without the need for replication inhibitors or DNA damage. We report here that the origin fork is processed in vivo to generate a regressed fork structure. Furthermore, origin fork regression supports two mechanisms of fork resolution that can potentially lead to fork reactivation. Fork regression generates both a site-specific double-stranded end (DSE) and a Holliday junction. Each of these DNA elements serves as a target for processing by the T4 ATPase/exonuclease complex [gene product (gp) 46/47] and Holliday junction-cleaving enzyme (EndoVII), respectively. In the absence of both gp46 and EndoVII, regressed origin forks are stabilized and persist throughout infection. In the presence of EndoVII, but not gp46, there is significantly less regressed origin fork accumulation apparently due to cleavage of the regressed fork Holliday junction. In the presence of gp46, but not EndoVII, regressed origin fork DSEs are processed by degradation of the DSE and a pathway that includes recombination proteins. Although both mechanisms can occur independently, they may normally function together as a single fork reactivation pathway.


Unintentional injury and its prevention in infant: knowledge and self-reported practices of main caregivers.
Unintentional injuries are the major cause of morbidity and mortality in infants. Prevention of unintentional injuries has been shown to be effective with education. Understanding the level of knowledge and practices of caregivers in infant safety would be useful to identify gaps for improvement. A cross-sectional study was conducted in an urban government health clinic in Malaysia among main caregivers of infants aged 11 to 15 months. Face-to-face interviews were conducted using a semi-structured self-designed questionnaire. Responses to the items were categorised by the percentage of correct answers: poor (<50%), moderate (50% - 70%) and good (>70%). A total of 403 caregivers participated in the study. Of the 21 items in the questionnaire on knowledge, 19 had good-to-moderate responses and two had poor responses. The two items on knowledge with poor responses were on the use of infant walkers (26.8%) and allowing infants on motorcycles as pillion riders (27.3%). Self-reported practice of infant safety was poor. None of the participants followed all 19 safety practices measured. Eight (42.1%) items on self-reported practices had poor responses. The worst three of these were on the use of baby cots (16.4%), avoiding the use of infant walkers (23.8%) and putting infants to sleep in the supine position (25.6%). Better knowledge was associated with self-reported safety practices in infants (p < 0.05). However, knowledge did not correspond to correct practice, particularly on the use of baby cots, infant walkers and sarong cradles. Main caregivers' knowledge on infant safety was good but self-reported practice was poor. Further research in the future is required to identify interventions that target these potentially harmful practices.


Tumor microenvironment as a metapopulation model: The effects of angiogenesis, emigration and treatment modalities.
Tumors consist of heterogeneous cell subpopulations that may develop differing phenotypes, such as increased cell growth, metastatic potential and treatment sensitivity or resistance. To study the dynamics of cancer development at a single-cell level, we model the tumor microenvironment as a metapopulation, in which habitat patches correspond to possible sites for cell subpopulations. Cancer cells may emigrate into dispersal pool (e.g. circulation system) and spread to new sites (i.e. metastatic disease). In the patches, cells divide and new variants may arise, possibly leading into an invasion provided the aberration promotes the cell growth. To study such adaptive landscape of cancer ecosystem, we consider various evolutionary strategies (phenotypes), such as emigration and angiogenesis, which are important determinants during early stages of tumor development. We use the metapopulation fitness of new variants to investigate how these strategies evolve through natural selection and disease progression. We further study various treatment effects and investigate how different therapy regimens affect the evolution of the cell populations. These aspects are relevant, for example, when examining the dynamic process of a benign tumor becoming cancerous, and what is the best treatment strategy during the early stages of cancer development. It is shown that positive angiogenesis promotes cancer cell growth in the absence of anti-angiogenic treatment, and that the anti-angiogenic treatment reduces the need of cytotoxic treatment when used in a combination. Interestingly, the model predicts that treatment resistance might become a favorable quality to cancer cells when the anti-angiogenic treatment is intensive enough. Thus, the optimal treatment dosage should remain below a patient-specific level to avoid treatment resistance.


Exploring the acute affective responses to resistance training: A comparison of the predetermined and the estimated repetitions to failure approaches.
In resistance-training (RT), the number of repetitions is traditionally prescribed using a predetermined approach (e.g., three sets of 10 repetitions). An emerging alternative is the estimated repetitions to failure (ERF) approach (e.g., terminating sets two repetitions from failure). Despite the importance of affective responses experienced during RT, a comparison between the two approaches on such outcomes is lacking. Twenty women (age range: 23-45 years) without RT experience completed estimated one repetition maximum (RM) tests in four exercises. In the next two counterbalanced sessions, participants performed the exercises using 70%1RM. Participants completed ten repetitions in all three sets (predetermined condition) or terminated the sets when perceived to be two repetitions away from task-failure (ERF condition). Primary outcomes were affective-valence, enjoyment, and approach-preference and secondary outcomes were repetition-numbers completed in each exercise. We observed trivial differences in the subjective measures and an approximately even approach-preference split. Under the ERF condition, we observed greater variability in repetition-numbers between participants and across exercises. Specifically, the mean number of repetitions was slightly lower in the chest-press, knee-extension, and lat-pulldown (~1 repetition) but considerably higher in the leg-press (17 vs. 10, p<0.01). Both approaches led to comparable affective responses and to an approximately even approach preference. Hence, prior to prescribing either approach, coaches should consider trainee's preferences. Moreover, under the ERF condition participants completed a dissimilar number of repetitions across exercises while presumably reaching a similar proximity to task-failure. This finding suggests that ERF allows for better effort regulation between exercises.


Nonylphenol diethoxylate inhibits apoptosis induced in PC12 cells.
Nonylphenol and short-chain nonylphenol ethoxylates such as NP2 EO are present in aquatic environment as wastewater contaminants, and their toxic effects on aquatic species have been reported. Apoptosis has been shown to be induced by serum deprivation or copper treatment. To understand the toxicity of nonylphenol diethoxylate, we investigated the effects of NP2 EO on apoptosis induced by serum deprivation and copper by using PC12 cell system. Nonylphenol diethoxylate itself showed no toxicity and recovered cell viability from apoptosis. In addition, nonylphenol diethoxylate decreased DNA fragmentation caused by apoptosis in PC12 cells. This phenomenon was confirmed after treating apoptotic PC12 cells with nonylphenol diethoxylate, whereas the cytochrome c release into the cytosol decreased as compared to that in apoptotic cells not treated with nonylphenol diethoxylates. Furthermore, Bax contents in apoptotic cells were reduced after exposure to nonylphenol diethoxylate. Thus, nonylphenol diethoxylate has the opposite effect on apoptosis in PC12 cells compared to nonylphenol, which enhances apoptosis induced by serum deprivation. The difference in structure of the two compounds is hypothesized to be responsible for this phenomenon. These results indicated that nonylphenol diethoxylate has capability to affect cell differentiation and development and has potentially harmful effect on organisms because of its unexpected impact on apoptosis. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1389-1398, 2016.


A comparative study of the five-repetition sit-to-stand test and the 30-second sit-to-stand test to assess exercise tolerance in COPD patients.
The sit-to-stand test (STST) has been used to evaluate the exercise tolerance of patients with COPD. However, mutual comparisons to predict poor exercise tolerance have been hindered by the variety of STST modes used in previous studies, which also did not consider patients' subjective perceptions of different STST modes. Our aim was to compare the five-repetition sit-to-stand test (5STS) with the 30-second sit-to-stand test (30STS) for predicting poor performance in the six-minute walking test and to evaluate patients' subjective perceptions to determine the optimal mode for clinical practice. Patients with stable COPD performed 5STS, 30STS and the 6MWT and then evaluated their feelings about the two STST modes by Borg dyspnea score and a questionnaire. Moreover, we collected data through the pulmonary function test, mMRC dyspnea score, COPD assessment test and quadriceps muscle strength (QMS). A receiver operating characteristic curve analysis of the 5STS and 30STS results was used to predict 6-minute walk distance (6MWD) <350 m. The final analysis included 128 patients. Similar moderate correlations were observed between 6MWT and 5STS (r=-0.508, P<0.001) and between 6MWT and 30STS (r=0.528, P<0.001), and there were similar correlations between QMS and 5STS (r=-0.401, P<0.001) and between QMS and 30STS (r=0.398, P<0.001). The 5STS and 30STS score cutoffs produced sensitivity, specificity and positive and negative predictive values of 76.0%, 62.8%, 56.7% and 80.3% (5STS) and 62.0%, 75.0%, 62.0% and 75.0% (30STS), respectively, for predicting poor 6MWT performance. The 5STS exhibited obvious superiority in terms of the completion rate and the subjective feelings of the participants. As a primary screening test for predicting poor 6MWD, the 5STS is similar to the 30STS in terms of sensitivity and specificity, but the 5STS has a better patient experience.


Calcium waves along the cleavage furrows in cleavage-stage Xenopus embryos and its inhibition by heparin.
Calcium signaling is known to be associated with cytokinesis; however, the detailed spatio-temporal pattern of calcium dynamics has remained unclear. We have studied changes of intracellular free calcium in cleavage-stage Xenopus embryos using fluorescent calcium indicator dyes, mainly Calcium Green-1. Cleavage formation was followed by calcium transients that localized to cleavage furrows and propagated along the furrows as calcium waves. The calcium transients at the cleavage furrows were observed at each cleavage furrow at least until blastula stage. The velocity of the calcium waves at the first cleavage furrow was approximately 3 microns/s, which was much slower than that associated with fertilization/egg activation. These calcium waves traveled only along the cleavage furrows and not in the direction orthogonal to the furrows. These observations imply that there exists an intracellular calcium-releasing activity specifically associated with cleavage furrows. The calcium waves occurred in the absence of extracellular calcium and were inhibited in embryos injected with heparin an inositol 1,4,5-trisphosphate (InsP3) receptor antagonist. These results suggest that InsP3 receptor-mediated calcium mobilization plays an essential role in calcium wave formation at the cleavage furrows.


Oscillations induced by different timescales in signal transduction modules regulated by slowly evolving protein-protein interactions.
The dynamics induced by the existence of different timescales in a system is explored, in the context of a model composed of activation and signalling modules regulated by a slowly evolving process, such as some particular protein-protein interactions or genetic-like dynamics. It is shown that slowly varying regulation patterns can induce rapid changes in the steady states of the (fast varying) signal transduction pathway, and lead to sustained oscillations. These results are illustrated by a reduced model of the Cdc2-cyclin B cell cycle oscillator. Using available experimental data, parameters of the model are estimated and found to agree with the requirements for a mechanism for oscillatory behaviour arising from coupling fast and slow processes.


Ulcer and gastritis.
Recent progress in the area of ulcer and gastritis is still dominated by findings and reports on Helicobacter pylori and nonsteroidal anti-inflammatory drugs, which in turn are the two major causes of peptic ulcers. Although the prevalence of H. pylori is declining in most developed countries, it is still contributing to a significant proportion of peptic ulcers globally. The interrelationship of H. pylori gastritis in patients with gastroesophageal reflux has become more apparent. H. pylori-induced gastric body gastritis is associated with reduced acid production, and thus with reduced reflux and esophagitis. The controversies regarding the interactions between H. pylori and NSAIDs have still not been settled. With the availability of the new COX-2-specific inhibitors, the current scenario of NSAID-related gastroduodenal complications will certainly change. Short-term usage of these agents has significantly reduced the incidence of endoscopic ulcers, but the benefits in terms of clinical outcomes, such as bleeding or perforation, remain to be determined. This review summarizes the recent literature on peptic ulcer and gastritis.


You are not always what you eat-Fatty acid bioconversion and lipid homeostasis in the larvae of the sand mason worm Lanice conchilega.
The meroplanktonic larvae of benthic organisms are an important seasonal component of the zooplankton in temperate coastal waters. The larvae of the reef-building polychaete Lanice conchilega contribute up to 15% of the summer zooplankton biomass in the North Sea. Despite their importance for reef maintenance (which positively affects the benthic community), little is known about the trophic ecology of this meroplanktonic larva. Qualitative and quantitative estimates of carbon (C) transfer between trophic levels and of fatty acid (FA)-specific assimilation, biosynthesis, and bioconversion can be obtained by compound-specific stable isotope analysis of FA. The present work tested the hypothesis that the concept of fatty acid trophic markers (FATM), widely used for studies on holoplankton with intermediate to high lipid contents, is also applicable to lipid-poor organisms such as meroplanktonic larvae. The incorporation of isotopically-enriched dietary C by L. conchilega larvae was traced, and lipid assimilation did not follow FA-specific relative availabilities in the diet. Furthermore, FAs that were unavailable in the diet, such as 22:5(n-3), were recorded in L. conchilega, suggesting their bioconversion by the larvae. The results indicate that L. conchilega larvae preferentially assimilate certain FAs and regulate their FA composition (lipid homeostasis) independently of that of their diet. Their quasi-homeostatic response to dietary FA availability could imply that the concept of FATM has limited application in lipid-poor organisms such as L. conchilega larvae.


Nonsurgical modalities to enhance healing and care of soft tissue wounds.
The rapidly aging population and patients with multiple concomitant pathologies present an increasing population of patients with nonhealing and problem wounds causing an unwelcome challenge for all health care providers. Many of these patients are not surgical candidates, or surgical procedures have failed to close their wounds. These wounds are particularly worrisome when an orthopaedic component is included, since bone and hardware must be covered as quickly as possible to prevent infection and even worse complications. We present a brief overview of several nonsurgical modalities that may be used to heal soft tissue wounds completely or to prepare the wound so a smaller surgical intervention may be done with greater chance for success. We include exogenous application of growth factors, cultured keratinocyte grafts, electrical stimulation, hyperbaric oxygen, and a vacuum-assisted closure system (V.A.C.).


Evolution of leaf form correlates with tropical-temperate transitions in Viburnum (Adoxaceae).
Strong latitudinal patterns in leaf form are well documented in floristic comparisons and palaeobotanical studies. However, there is little agreement about their functional significance; in fact, it is still unknown to what degree these patterns were generated by repeated evolutionary adaptation. We analysed leaf form in the woody angiosperm clade Viburnum (Adoxaceae) and document evolutionarily correlated shifts in leafing habit, leaf margin morphology, leaf shape and climate. Multiple independent shifts between tropical and temperate forest habitats have repeatedly been accompanied by a change between evergreen, elliptical leaves with entire margins and deciduous, more rounded leaves with toothed or lobed margins. These consistent shifts in Viburnum support repeated evolutionary adaptation as a major determinant of the global correlation between leaf form and mean annual temperature. Our results provide a new theoretical grounding for the inference of past climates using fossil leaf assemblages.


Application of a Recombinant Three-Factor Chromogenic Reagent, PyroSmart, for Bacterial Endotoxins Test Filed in the Pharmacopeias.
Assays using lysate reagents prepared from horseshoe crab hemocyte extract (limulus amoebocyte lysate, LAL) are commonly and widely used to detect and measure endotoxin in parenteral drugs and medical devices. However, lysate reagents suffer from lot-to-lot variations leading to possible fluctuations in testing. Also, this continued usage of lysate reagents leads to the possible decline of the horseshoe crab population. Recently, a new recombinant chromogenic reagent, PyroSmart, consisting of three recombinant factors was introduced to the market. There are now three recombinant products; two with recombinant factor C reagents and PyroSmart with the complete recombinant LAL system. We evaluated the applicability of the reagent to the harmonized bacterial endotoxins test in the United States, European and Japanese pharmacopeias. The recombinant product showed equivalent potency of thirteen endotoxins from different bacterial strains to conventional chromogenic lysate reagents as long as their assay modes are identical. All analytical characteristics or assay parameters of the reagent satisfied the acceptance criteria which are set for the use for the bacterial endotoxins test filed in the pharmacopeias. All of 109 parenteral drugs tested can be measured with PyroSmart within respective maximum allowable dilutions. The lot-to-lot variation in recovery of endotoxin added in the parenteral drugs for PyroSmart was equal to or less than those of six limulus lysate reagents. In conclusion, the present study suggests that the recombinant reagent, PyroSmart, provide a good alternative to the LAL reagents with better lot-to-lot variation.


The UBIRIS.v2: a database of visible wavelength iris images captured on-the-move and at-a-distance.
The iris is regarded as one of the most useful traits for biometric recognition and the dissemination of nationwide iris-based recognition systems is imminent. However, currently deployed systems rely on heavy imaging constraints to capture near infrared images with enough quality. Also, all of the publicly available iris image databases contain data correspondent to such imaging constraints and therefore are exclusively suitable to evaluate methods thought to operate on these type of environments. The main purpose of this paper is to announce the availability of the UBIRIS.v2 database, a multisession iris images database which singularly contains data captured in the visible wavelength, at-a-distance (between four and eight meters) and on on-the-move. This database is freely available for researchers concerned about visible wavelength iris recognition and will be useful in accessing the feasibility and specifying the constraints of this type of biometric recognition.


Memory Disorders Related to Hippocampal Function: The Interest of 5-HT4Rs Targeting.
The hippocampus has long been considered as a key structure for memory processes. Multilevel alterations of hippocampal function have been identified as a common denominator of memory impairments in a number of psychiatric and neurodegenerative diseases. For many years, the glutamatergic and cholinergic systems have been the main targets of therapeutic treatments against these symptoms. However, the high rate of drug development failures has left memory impairments on the sideline of current therapeutic strategies. This underscores the urgent need to focus on new therapeutic targets for memory disorders, such as type 4 serotonin receptors (5-HT4Rs). Ever since the discovery of their expression in the hippocampus, 5-HT4Rs have gained growing interest for potential use in the treatment of learning and memory impairments. To date, much of the researched information gathered by scientists from both animal models and humans converge on pro-mnesic and anti-amnesic properties of 5-HT4Rs activation, although the mechanisms at work require more work to be fully understood. This review addresses a fundamental, yet poorly understood set of evidence of the potential of 5-HT4Rs to re-establish or limit hippocampal alterations related to neurological diseases. Most importantly, the potential of 5-HT4Rs is translated by refining hypotheses regarding the benefits of their activation in memory disorders at the hippocampal level.


Impact of the COVID-19 pandemic on hospitalizations for acute coronary syndromes: a multinational study.
COVID-19 has challenged the health system organization requiring a fast reorganization of diagnostic/therapeutic pathways for patients affected by time-dependent diseases such as acute coronary syndromes (ACS). To describe ACS hospitalizations, management, and complication rate before and after the COVID-19 pandemic was declared. Ecological retrospective study. Methods: We analyzed aggregated epidemiological data of all patients > 18 years old admitted for ACS in twenty-nine hub cardiac centers from 17 Countries across 4 continents, from December 1st, 2019 to April 15th, 2020. Data from December 2018 to April 2019 were used as historical period. A significant overall trend for reduction in the weekly number of ACS hospitalizations was observed (20.2%; 95% confidence interval CI [1.6, 35.4] P = 0.04). The incidence rate reached a 54% reduction during the second week of April (incidence rate ratio: 0.46, 95% CI [0.36, 0.58]) and was also significant when compared to the same months in 2019 (March and April, respectively IRR: 0.56, 95%CI [0.48, 0.67]; IRR: 0.43, 95%CI [0.32, 0.58] p < 0.001). A significant increase in door-to-balloon, door-to-needle, and total ischemic time (p <0.04 for all) in STEMI patents were reported during pandemic period. Finally, the proportion of patients with mechanical complications was higher (1.98% vs. 0.98%; P = 0.006) whereas GRACE risk score was not different. Our results confirm that COVID-19 pandemic was associated with a significant decrease in ACS hospitalizations rate, an increase in total ischemic time and a higher rate of mechanical complications on a international scale.


The role of laparoscopy in the diagnosis and management of heterotopic pregnancies.
The object of this report is to discuss diagnosis and treatment of heterotopic pregnancies. Thirteen consecutive cases referred to our institution are reviewed. In 54% of cases the heterotopic pregnancy was asymptomatic. The ectopic pregnancy was visualized prior to surgery in 69% of the cases. The treatment was surgical in every case and performed laparoscopically in 77% of cases. Ten patients underwent salpingectomy and three salpingostomy. In all, 60% of intrauterine pregnancies that were viable at the time of diagnosis of the heterotopic pregnancy had a favourable outcome. Diagnosis of heterotopic pregnancy is difficult. Laparoscopy allows both diagnosis and treatment, and the outcome of the intrauterine pregnancy is comparable to that obtained with laparotomy.


Enantioselective induction of oxidative stress by permethrin in rat adrenal pheochromocytoma (PC12) cells.
Synthetic pyrethroid (SP) insecticides are chiral compounds with multiple asymmetric positions. Several recent studies have focused on the effect of enantioselectivity of SPs in acute aquatic toxicity, endocrine-disrupting activities, and immunotoxicity. However, the relevant molecular mechanisms are still unknown. The potential relationship between ecotoxicological effects and oxidative stress could contribute to SP-induced enantioselective cytotoxicity, but this requires further investigation. Therefore, this study was undertaken to evaluate the role of oxidative stress in enantiomer-specific permethrin (PM)-induced cytotoxicity in rat adrenal pheochromocytoma (PC12) cells. The study demonstrated that PM induced enantioselective oxidative stress and cytotoxicity. The reactive oxygen species (ROS) generation and lipid peroxidation production of malondialdehyde (MDA) were obviously increased, whereas the activity of antioxidant enzymes (superoxide dismutase [SOD] and catalase [CAT]) and glutathione (GSH) content had declined after exposure in 1R-trans-PM at a concentration of 30 mg/L. Meanwhile, the result of the cytotoxicity assay showed a clear, dose-dependent growth-inhibition effect of PM in an enantioselective manner. The most toxic effect on PC12 cells was shown by 1R-trans-PM and was approximately 1.6 times higher than that with 1S-cis-PM, which exhibited only a slightly toxic effect at a concentration of 20 mg/L. These results suggested that PM exhibited significant enantioselectivity in oxidative stress, which may be one of the initial events in PM-induced enantioselective cytotoxicity. The present study also improved understanding of enantiomer-specific, SP-induced cytotoxicity. The enantioselectivity should be taken into consideration when assessing ecological effects and development of new chiral pesticides.


Plasmodium falciparum: association with erythrocytic superoxide dismutase.
Levels of superoxide dismutase (SOD) activity and its properties in Plasmodium falciparum-infected erythrocytes, isolated parasites, and noninfected erythrocytes were studied. A higher specific activity was found in P. falciparum-infected erythrocytes compared to noninfected erythrocytes, resulting from the lower protein content of infected cells and not enzyme synthesis by the parasite, as the superoxide dismutase activity expressed per number of cells was decreased. Superoxide dismutase from noninfected erythrocytes and isolated P. falciparum parasites showed similar sensitivities to various inhibitors and had identical molecular weights and electrophoretic mobilities. These results support the hypothesis of uptake and use of the erythrocytic SOD enzyme by the parasite as a possible mechanism of defense against oxidative stress.


Distinct functions of histone H3, lysine 4 methyltransferases in normal and malignant hematopoiesis.
Histone H3, lysine 4 (H3K4) methylation is one chromatin modification that defines distinct regulatory states of euchromatin. Mammals express six main histone methyltransferase (HMT) enzymes that modify H3K4 by monomethylation, dimethylation or trimethylation. Recent studies examine roles of some of these HMTs and their cofactors in hematopoiesis and leukemia. We discuss these emerging studies together with prior embryonic stem data, revealing how these enzymes function. Murine models have been employed to conditionally or constitutively knockout HMTs (MLL1/KMT2A, MLL2/KMT2B, MLL3/KMT2C, MLL4/KMT2D, SETD1A/KMT2F and SETD1B/KMT2G) as well as specific domains or partners of these enzymes in normal hematopoietic populations and in the context of hematologic malignancies. These studies demonstrate that global or gene-specific changes in H3K4 modification levels can be attributed to particular enzymes in particular tissues. Loss-of-function studies indicate largely nonoverlapping roles of the six H3K4 HMTs. These roles are not all necessarily due to differences in enzymatic activity and are not always accompanied by large global changes in histone modification. Both gain-of-function and loss-of-function mutations in hematologic malignancy are restricted to MLL1 and MLL3/MLL4, but emerging data indicate that SETD1A/SETD1B and MLL2 can be critical in leukemia as well.


Spatial variation and clustering of anaemia prevalence in school-aged children in Western Kenya.
Anaemia surveillance has overlooked school-aged children (SAC), hence information on this age group is scarce. This study examined the spatial variation of anaemia prevalence among SAC (5-14 years) in western Kenya, a region associated with high malaria infection rates. A total of 8051 SAC were examined from 82 schools across eight counties in Western Kenya in February 2022. Haemoglobin (Hb) concentrations were assessed at the school and village level and anaemia defined as Hb<11.5g/dl for age 5-11yrs and Hb <12.0g/dl for 12-14yrs after adjusting for altitude. Moran's I analysis was used to measure spatial autocorrelation, and local clusters of anaemia were mapped using spatial scan statistics and local indices of spatial association (LISA). The prevalence of anaemia among SAC was 27.8%. The spatial variation of anaemia was non-random, with Global Moran's I 0.2 (p-value < 0.002). Two significant anaemia cluster windows were identified: Cluster 1 (LLR = 38.9, RR = 1.4, prevalence = 32.0%) and cluster 2 (LLR = 23.6, RR = 1.6, prevalence = 45.5%) at schools and cluster 1 (LLR = 41.3, RR = 1.4, prevalence = 33.3%) and cluster 2 (LLR = 24.5, RR = 1.6, prevalence = 36.8%) at villages. Additionally, LISA analysis identified ten school catchments as anaemia hotspots corresponding geographically to SatScan clusters. Anaemia in the SAC is a public health problem in the Western region of Kenya with some localised areas presenting greater risk relative to others. Increasing coverage of interventions, geographically targeting the prevention of anaemia in the SAC, including malaria, is required to alleviate the burden among children attending school in Western Kenya.


Structural Basis for Targeting T:T Mismatch with Triaminotriazine-Acridine Conjugate Induces a U-Shaped Head-to-Head Four-Way Junction in CTG Repeat DNA.
The potent DNA-binding compound triaminotriazine-acridine conjugate (Z1) functions by targeting T:T mismatches in CTG trinucleotide repeats that are responsible for causing neurological diseases such as myotonic dystrophy type 1, but its binding mechanism remains unclear. We solved a crystal structure of Z1 in a complex with DNA containing three consecutive CTG repeats with three T:T mismatches. Crystallographic studies revealed that direct intercalation of two Z1 molecules at both ends of the CTG repeat induces thymine base flipping and DNA backbone deformation to form a four-way junction. The core of the complex unexpectedly adopts a U-shaped head-to-head topology to form a crossover of each chain at the junction site. The crossover junction is held together by two stacked G:C pairs at the central core that rotate with respect to each other in an X-shape to form two nonplanar minor-groove-aligned G·C·G·C tetrads. Two stacked G:C pairs on both sides of the center core are involved in the formation of pseudo-continuous duplex DNA. Four metal-mediated base pairs are observed between the N7 atoms of G and CoII, an interaction that strongly preserves the central junction site. Beyond revealing a new type of ligand-induced, four-way junction, these observations enhance our understanding of the specific supramolecular chemistry of Z1 that is essential for the formation of a noncanonical DNA superstructure. The structural features described here serve as a foundation for the design of new sequence-specific ligands targeting mismatches in the repeat-associated structures.


Adrenal scintigraphy with 131-I-adosterol.
A new adrenal scanning agent, Adosterol (131I-6beta-iodomethyl-19-norcholest-5(10)-en-3beta-ol) has been evaluated in three female patients with hypercorticism. In one case, characterised by recurrent adrenocortical hyperactivity after bilateral adrenalectomy, a remnant of adrenal tissue was detected. In two other cases, the presence of an adrenal tumour could be excluded. The scan made on the 6th day after the administration of Adosterol was found to be superior in quality to that made on the 13th day.


[Comparative study of five different methods for grouping beta-haemolytic streptococci (author's transl)].
161 strains of beta-haemolytic streptococci, that had been isolated from patients' material, and 7 reference strains were group]ed with the techniques of precipitation, direct immunofluorescence, co-agglutination (Phadebact Streptococcus Test), latex agglutination (Streptex), and biochemical tests (Api-Strep). From the reference strains one group G strain was incorrectly identified with the Api-Strep. The direct immunofluorescence test yielded a result in agreement with the precipitation method in 94% of the freshly isolated strains. Co-agglutination and latex agglutination gave corresponding results with the precipitation method in 99%, taking into consideration that the first method doesn't apply to groups D and F. The biochemical determination with Api-Strep gave correct results with all strains of groups A and B, but did not do so with some group G and most of the group C streptococcal strains. Therefore especially co-agglutination and latex agglutination can be recommended for routine diagnostic purposes as reliable, rapid, and simple methods.


[Endoscopic treatment of intracranial lesions. Apropos of 8 cases].
The authors describe their initial experience involving endoscopic techniques used in the treatment of eight patients with varied brain lesions. Two tumours and a colloid cyst of the third ventricle, two paraventricular symptomatic cysts, one arachnoid cyst, one chronic intracerebral hematoma and one case of free catheter extraction in the lateral ventricle. The common characteristics of all these lesions were their liquid character and/or intraventricular location. Rigid endoscopes were used, with 9 and 6 mm work channels and independent aspiration and irrigation systems. The optic elbow makes direct visualization and adaptation to the TV monitor possible. The endoscope was manually directed at the lesion through a 12 mm burr hole in five cases and by means of a stereotactic frame in three cases. Biopsies were obtained in six cases. In three cases a communication between the cyst and the ventricular system or adjacent cisterns was established and in two cases vaporization of the lesion using a CO2 laser was performed. Tolerance to the endoscopic procedures was good in all cases. In the authors' opinion, endoscopic techniques constitute a low risk method, midway between stereotactic techniques and the microsurgical approach, and are especially indicated in the treatment of cystic brain lesions or those situated in or near the ventricular system. Their most notable advantages are: providing direct visualization of the lesion without interference from the instruments in the operating field; non-repercussion of the spatial changes derived from the liquid outlet and possibility of lesion coagulation and manipulation. Their disadvantages, related to the use of very long instruments and from a reduced work channel, must be overcome by specialized training.


Predictors of patrol officer interest in cybercrime training and investigation in selected United States police departments.
Cybercrime has created substantial challenges for law enforcement, particularly at the local level. Most scholars and police administrators believe that patrol officers need to become more effective first responders to cybercrime calls. The evidence illustrates, however, that many patrol officers are neither adequately prepared nor strongly interested in taking an active role in addressing cybercrime at the local level. This study, therefore, examined the factors that predicted patrol officer interest in cybercrime training and investigations in two southeastern U.S. cities. The study specifically examined the relationship between demographics, cybercrime exposure, computer training, computer proficiency, Internet and cybercrime perceptions, and views on policing cybercrime with officer interest in cybercrime investigation training and conducting cybercrime investigations in the future. Officer views on policing cybercrime, particularly whether they valued cybercrime investigations and believed that cybercrime would dramatically change policing, along with their computer skills, were the strongest predictors of interest in cybercrime efforts. Officers who had received previous computer training were less interested in additional training and conducting investigations. These findings support the argument that more command and departmental meetings focusing on the value of investigating these types of crime need to be held in order to increase officer interest.


The origin of regenerating mesothelium: a historical perspective.
Bichat first described the mesothelium in 1827 but despite its early discovery, it has only been in recent years that its importance both in health and disease has been realised. One area still poorly understood is that of the mechanisms regulating mesothelial repair. Mesothelial cells are derived from the mesoderm but express many epithelial characteristics. However, mesothelium does not heal in the same way as other epithelial-like cells. Epithelium heals by centripetal migration, with cells at the edge of the wound proliferating and migrating into the injured area. Hertzler in 1919 noted that both large and small peritoneal injuries healed within the same time frame, concluding that the mesothelium could not heal solely by centripetal migration. The exact mechanisms involved in mesothelial regeneration following injury are controversial with a number of proposals suggested to explain the origin of the regenerating cells. This review will examine these proposals and give some insights into the likely mechanisms involved.


Student-, classroom-, and school-level risk factors for victimization.
The purpose of this study was to simultaneously investigate student-, classroom-, and school-level risk factors for victimization. Both peer nominations and students' self-reports of victimization were utilized. The sample consisted of 6731 Finnish elementary school students (3386 girls and 3345 boys) nested in 358 classrooms in 74 schools. The participants were from Grades 3, 4, and 5 (mean age 11years). The results of multilevel analyses indicated that there was considerable variability in, and distinctive risk factors associated with, both peer- and self-reported victimization at all the three levels investigated. Social anxiety and peer rejection synergistically predicted victimization at the student level. At the classroom level, negative social outcome expectations of defending the victim were associated with an increased risk of a student being bullied. Victimization was also common in classrooms and schools where students perceived their teachers to have less disapproving attitudes toward bullying. Furthermore, the effects of the student-level predictors were found to vary across classrooms, and classroom size moderated the effects of social anxiety and peer rejection on victimization. By identifying the risk factors at the multiple levels, and looking into cross-level interactions among these factors, research can help to target interventions at the key ecological factors contributing to victimization, making it possible to maximize the effectiveness of interventions.


Effects of isovalerate supplements on morphology and functional gene expression of rumen mucosa in pre- and post-weaning dairy calves.
Isovalerate supplements could stimulate rumen development by improving morphology and function of rumen mucosa, and then promote the growth of calves. This study was done to evaluate the effects of isovalerate supplements on morphology and functional gene expression of rumen mucosa in dairy calves. In total, 48 Chinese Holstein male calves with 15 days of age and 45.1±0.36 kg of BW were randomly assigned to four groups. The treatments were: control, low-isovalerate, moderate-isovalerate and high-isovalerate with 0, 3, 6 and 9 g isovalerate per calf per day, respectively. Supplementary isovalerate was hand-mixed into milk in pre-weaning calves and into concentrate portion in post-weaning calves. The study consisted of a 15-day-adaptation period and a 60-day-sampling period. Calves were weaned at 60 days of age. Three calves were slaughtered from each of the four treatments at 30, 60 and 90 days of age. The weight of body and stomach were measured, samples of ruminal tissues and blood were analyzed. Total stomach weight, total stomach to BW ratio, rumen wall and keratinized layer thickness, serum growth hormone and IGF-1 for both pre- and post-weaning calves increased linearly with increasing isovalerate supplements. Rumen to total stomach weight ratio, the length and width of rumen papillae, and serum β-hydroxybutyrate increased linearly for post-weaning calves. However, abomasum weight to total stomach weight ratio decreased linearly for both pre- and post-weaning calves. The relative messenger RNA expression for growth hormone receptor, IGF-1 receptor and 3-hydroxy-3-methylglutaryl-CoA synthase 1 in rumen mucosa increased linearly for post-weaning calves. Our results suggested that isovalerate supplements promoted rumen development in a dose-dependent manner. The optimum dose was 6.0 g isovalerate per calf per day.


Men who self-harm-A scoping review of a complex phenomenon.
To understand and care for men who self-harm, it is important that healthcare professionals have understanding of how and why men self-harm, men's experiences of self-harm and what can be done to hinder or prevent self-harm. The aim of this study was to synthesize the existing knowledge on men who self-harm, with a special emphasis on background, self-harming methods, experiences and reported therapeutic interventions and/or care approaches. Scoping review of internationally published and grey literature, based on a methodological framework by Arksey and O'Malley. Systematic electronic database searches were conducted in CINAHL, MEDLINE (Ovid) and PsycINFO. From a total of 684 studies found, 24 studies met the inclusion criteria: full-text, published in English, peer-reviewed studies and grey literature including a focus on men who self-harm, men aged between 18 and 65 years, and published between 2010 and 2019. Men's self-harm was understood as being related to mental disorders, a means of affect regulation, a loss of self-control, and a means of interpersonal communication. Self-harm can be a positive or negative experience, and there is a wide variety in the methods that men use to self-harm: sharp objects, injection, ingestion, without aids or riskful behaviour. Few studies reported on therapeutic interventions and/or care approaches for men who self-harm. Men's self-harm should be understood as a complex, socially and culturally conditioned phenomenon and studied from a multitude of perspectives. This scoping review concludes that self-harm among men should be understood as a complex, socially and culturally conditioned phenomenon. To empower men and support their recovery from self-harm, a person-centred approach should be incorporated into research on the subject and practice.


In vitro detection of bacterial contamination in platelet concentrates by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: a preliminary study.
Platelet concentrates are at risk of transfusion-related sepsis. The microbial detection methods currently available have reached their limits, as they do not completely prevent transfusion-related bacterial contamination.The aim of this study was to develop a new strategy to detect the risk of platelet transfusion-related bacterial contamination using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In vitro, platelet concentrates were seeded with known concentrations of bacterial strains. Protein mass profiles were acquired by using a Microflex MALDI-TOF instrument. Dedicated 'Platelet' software was used as a spectrum subtraction tool to reveal specific peaks caused by the presence of pathogens in samples. The MALDI-TOF spectra of platelets were characterized and the reproducibility over time, regardless of the blood donor, was demonstrated with a positive predictive value of 100 %. In addition, the negative predictive value of the total number of specific peaks to predict contamination was 100 %. Detecting bacteria in platelet concentrates using the MALDI-TOF approach and analysing spectra with the Platelet software present the advantage of combining the precocity of results and sufficient sensitivity (10 c.f.u. ml-1). Further research will be conducted to compare this novel method with the current conventional method in order to validate our results, the objective being to reduce the risk of platelet transfusion-related bacterial contamination.


Complications of intra- and peri-articular steroid injections.
A prospective study was conducted to investigate the possible complications associated with intra- and peri-articular steroid injections. Data are presented on 1147 injections performed on 672 patients out of a total of 688 consecutive patients attending an orthopaedic outpatient setting. It was found to be a safe procedure, with a very low complication rate, if performed while taking adequate precautions. This should encourage general practitioners to offer these injections in their surgeries.


Structural intermediates in the assembly of taxoid-induced microtubules and GDP-tubulin double rings: time-resolved X-ray scattering.
We have studied the self-association reactions of purified GDP-liganded tubulin into double rings and taxoid-induced microtubules, employing synchrotron time-resolved x-ray solution scattering. The experimental scattering profiles have been interpreted by reference to the known scattering profiles to 3 nm resolution and to the low-resolution structures of the tubulin dimer, tubulin double rings, and microtubules, and by comparison with oligomer models and model mixtures. The time courses of the scattering bands corresponding to the different structural features were monitored during the assembly reactions under varying biochemical conditions. GDP-tubulin essentially stays as a dimer at low Mg(2+) ion activity, in either the absence or presence of taxoid. Upon addition of the divalent cations, it associates into either double-ring aggregates or taxoid-induced microtubules by different pathways. Both processes have the formation of small linear (short protofilament-like) tubulin oligomers in common. Tubulin double-ring aggregate formation, which is shown by x-ray scattering to be favored in the GDP- versus the GTP-liganded protein, can actually block microtubule assembly. The tubulin self-association leading to double rings, as determined by sedimentation velocity, is endothermic. The formation of the double-ring aggregates from oligomers, which involves additional intermolecular contacts, is exothermic, as shown by x-ray and light scattering. Microtubule assembly can be initiated from GDP-tubulin dimers or oligomers. Under fast polymerization conditions, after a short lag time, open taxoid-induced microtubular sheets have been clearly detected (monitored by the central scattering and the maximum corresponding to the J(n) Bessel function), which slowly close into microtubules (monitored by the appearance of their characteristic J(0), J(3), and J (n) - (3) Bessel function maxima). This provides direct evidence for the bidimensional assembly of taxoid-induced microtubule polymers in solution and argues against helical growth. The rate of microtubule formation was increased by the same factors known to enhance taxoid-induced microtubule stability. The results suggest that taxoids induce the accretion of the existing Mg(2+)-induced GDP-tubulin oligomers, thus forming small bidimensional polymers that are necessary to nucleate the microtubular sheets, possibly by binding to or modifying the lateral interaction sites between tubulin dimers.


Cloning and functional analysis of HpFAD2 and HpFAD3 genes encoding Δ12- and Δ15-fatty acid desaturases in Hansenula polymorpha.
Two fatty acid desaturase genes have been cloned: HpFAD2 and HpFAD3 encode Hansenula polymorpha Δ12-fatty acid desaturase (HpFad2) and Δ15-fatty acid desaturase (HpFad3), which are responsible for the production of linoleic acid (LA, C18:2, Δ9, Δ12) and α-linolenic acid (ALA, αC18:3, Δ9, Δ12, Δ15), respectively. The open reading frame of the HpFAD2 and HpFAD3 genes is 1215bp and 1239bp, encoding 405 and 413 amino acids, respectively. The putative amino acid sequences of HpFad2 and HpFad3 share more than 60% similarity and three conserved histidine-box motifs with other known yeast Fad homologs. Hpfad2Δ disruptant cannot produce C18:2 and αC18:3, while the deletion of HpFAD3 only causes the absence of αC18:3. Heterologous expression of either the HpFAD2 or the HpFAD3 gene in Saccharomyces cerevisiae resulted in the presence of C18:2 and αC18:3 when the C18:2 precursor was added. Taken together, these observations indicate that HpFAD2 and HpFAD3 indeed encode Δ12- and Δ15-fatty acid desaturases that function as the only ones responsible for desaturation of oleic acid (C18:1) and linoleic acid (C18:2), respectively, in H. polymorpha. Because a Fatty Acid Regulated (FAR) region and a Low Oxygen Response Element (LORE), which are responsible for regulation of a Δ9-fatty acid desaturase gene (ScOLE1) in S. cerevisiae, are present in the upstream regions of both genes, we investigated whether the transcriptional levels of HpFAD2 and HpFAD3 are affected by supplementation with nutrient unsaturated fatty acids or by low oxygen conditions. Whereas both genes were up-regulated under low oxygen conditions, only HpFAD3 transcription was repressed by an excess of C18:1, C18:2 and C18:3, while the HpFAD2 transcript level did not significantly change. These observations indicate that HpFAD2 expression is not controlled at the transcriptional level by fatty acids even though it contains a FAR-like region. This study indicates that HpFAD2 may be regulated by post-transcriptional mechanisms, whereas HpFAD3 may be mainly controlled at a transcriptional level.


Operative treatment of osteochondral lesions of the talus.
➤ Osteochondral lesions of the talus are common injuries in recreational and professional athletes, with up to 50% of acute ankle sprains and fractures developing some form of chondral injury. Surgical treatment paradigms aim to restore the articular surface with a repair tissue similar to native cartilage and to provide long-term symptomatic relief.➤ Arthroscopic bone-marrow stimulation techniques, such as microfracture and drilling, perforate the subchondral plate with multiple openings to recruit mesenchymal stem cells from the underlying bone marrow to stimulate the differentiation of fibrocartilaginous repair tissue in the defect site. The ability of fibrocartilage to withstand mechanical loading and protect the subchondral bone over time is a concern.➤ Autologous osteochondral transplantation techniques replace the defect with a tubular unit of viable hyaline cartilage and bone from a donor site in the ipsilateral knee. In rare cases, a graft can also be harvested from the ipsilateral talus or contralateral knee. The limitations of donor site morbidity and the potential need for an osteotomy about the ankle should be considered. Some anterior or far posterior talar lesions can be accessed without arthrotomy or with a plafondplasty.➤ Osteochondral allograft transplantation allows an osteochondral lesion with a large surface area to be replaced with a single unit of viable articular cartilage and subchondral bone from a donor that is matched to size, shape, and surface curvature. The best available evidence suggests that this procedure should be limited to large-volume cystic lesions or salvage procedures.➤ Autologous chondrocyte implantation techniques require a two-stage procedure, the first for chondrocyte harvest and the second for implantation in a periosteum-covered or matrix-induced form after in vivo culture expansion. Theoretically, the transplantation of chondrocyte-like cells into the defect will result in hyaline-like repair tissue.