Diagnostic utility of galectin-3 in thyroid cancer.
Galectin-3 (Gal-3), which has received significant recent attention for its utility as a diagnostic marker for thyroid cancer, represents the most well-studied molecular candidate for thyroid cancer diagnosis. Gal-3 is a protein that binds to beta-galactosidase residues on cell surface glycoproteins and has also been identified in the cytoplasmic and nuclear compartment. This marker has been implicated in regulation of normal cellular proliferation and apoptosis, as well as malignant transformation and the metastasis of cancer cells. We here present a mechanistic review of Gal-3 and its role in cancer development and progression. Gal-3 expression studies in thyroid tissue and cytologic tumor specimens and their methodological considerations are also discussed in this article. Despite great variance in their methodology, the majority of immunohistochemical studies found that Gal-3 was differentially expressed in thyroid carcinoma compared with benign and normal thyroid specimens, suggesting that Gal-3 is a good diagnostic marker for thyroid cancer. Recent studies have also demonstrated improved methodological reliability. On the other hand, Gal-3 genomic expression studies have shown inconsistent results for diagnostic utility and are not recommended. Overall, the development of Gal-3 as a diagnostic marker for thyroid cancer represents a promising avenue for future study, and its clinical application could significantly reduce the number of diagnostic thyroid operations performed for cases of indeterminant fine needle aspiration biopsy cytology, and thus positively impact the current management of thyroid nodular disease.


Exploring the role of post-translational modifications in regulating α-synuclein interactions by studying the effects of phosphorylation on nanobody binding.
Intracellular deposits of α-synuclein in the form of Lewy bodies are major hallmarks of Parkinson's disease (PD) and a range of related neurodegenerative disorders. Post-translational modifications (PTMs) of α-synuclein are increasingly thought to be major modulators of its structure, function, degradation and toxicity. Among these PTMs, phosphorylation near the C-terminus at S129 has emerged as a dominant pathogenic modification as it is consistently observed to occur within the brain and cerebrospinal fluid (CSF) of post-mortem PD patients, and its level appears to correlate with disease progression. Phosphorylation at the neighboring tyrosine residue Y125 has also been shown to protect against α-synuclein toxicity in a Drosophila model of PD. In the present study we address the potential roles of C-terminal phosphorylation in modulating the interaction of α-synuclein with other protein partners, using a single domain antibody fragment (NbSyn87) that binds to the C-terminal region of α-synuclein with nanomolar affinity. The results reveal that phosphorylation at S129 has negligible effect on the binding affinity of NbSyn87 to α-synuclein while phosphorylation at Y125, only four residues away, decreases the binding affinity by a factor of 400. These findings show that, despite the fact that α-synuclein is intrinsically disordered in solution, selective phosphorylation can modulate significantly its interactions with other molecules and suggest how this particular form of modification could play a key role in regulating the normal and aberrant function of α-synuclein.


Transcriptome analysis of desmoplastic small round cell tumors identifies actionable therapeutic targets: a report from the Children's Oncology Group.
To further understand the molecular pathogenesis of desmoplastic small round cell tumor (DSRCT), a fatal malignancy occurring primarily in adolescent/young adult males, we used next-generation RNA sequencing to investigate the gene expression profiles intrinsic to this disease. RNA from DSRCT specimens obtained from the Children's Oncology Group was sequenced using the Illumina HiSeq 2000 system and subjected to bioinformatic analyses. Validation and functional studies included WT1 ChIP-seq, EWS-WT1 knockdown using JN-DSRCT-1 cells and immunohistochemistry. A panel of immune signature genes was also evaluated to identify possible immune therapeutic targets. Twelve of 14 tumor samples demonstrated presence of the diagnostic EWSR1-WT1 translocation and these 12 samples were used for the remainder of the analysis. RNA sequencing confirmed the lack of full-length WT1 in all fusion positive samples as well as the JN-DSRCT-1 cell line. ChIP-seq for WT1 showed significant overlap with genes found to be highly expressed, including IGF2 and FGFR4, which were both highly expressed and targets of the EWS-WT1 fusion protein. In addition, we identified CD200 and CD276 as potentially targetable immune checkpoints whose expression is independent of the EWS-WT1 fusion gene in cultured DSCRT cells. In conclusion, we identified IGF2, FGFR4, CD200, and CD276 as potential therapeutic targets with clinical relevance for patients with DSRCT.


A high-throughput screen for compounds that inhibit aggregation of the Alzheimer's peptide.
Aggregation of the Alzheimer's peptide Abeta produces toxic multimeric species that play a key role in the development of Alzheimer's disease. Compounds that inhibit this aggregation may prove useful as therapeutic agents for the prevention or treatment of Alzheimer's disease. Although aggregation inhibitors may already exist in combinatorial libraries, finding these compounds in a cost-effective high-throughput manner poses an enormous challenge. To meet this challenge, we have developed a novel high-throughput screen capable of isolating inhibitors of Abeta aggregation from large libraries of inactive candidates. The screen uses a fusion of Abeta42 to GFP. In the absence of inhibition, the rapid misfolding and aggregation of Abeta42 causes the entire fusion protein to misfold, thereby preventing fluorescence. Compounds that inhibit Abeta42 aggregation enable GFP to fold into its native structure and be identified by the resulting fluorescent signal. By implementing the screen on a pilot library of triazine derivatives, we have identified several putative inhibitors. One of the selected compounds was studied in detail by a series of biochemical and biophysical methods. These studies confirmed that the selected compound inhibits aggregation of synthetic Abeta42 peptide. The fluorescence-based method described here is rapid and inexpensive and can be used to screen large libraries for inhibitors of Abeta42 aggregation and/or amyloidogenesis.


Validation of the SETOC instrument -- Student evaluation of teaching in outpatient clinics.
There is a paucity of evaluation forms specifically developed and validated for outpatient settings. The purpose of this study was to develop and validate an instrument specifically for evaluating outpatient teaching, to provide reliable and valid ratings for individual and group feedback to faculty, and to identify outstanding teachers in that setting. By literature reviews and pilot studies at the Faculties of Health Sciences, McMaster University (Canada) and Aga Khan University (AKU-Pakistan), a 15-item instrument, Student Evaluation of Teaching in Outpatient Clinics (SETOC), was created with five subscales: "Establishing Learning-Milieu, Clinical-Teaching, General-Teaching, Clinical-Competence, and Global-Rating." Seven-point Likert-type rating scales were used. Students also nominated three "best" outpatient teachers. 87 faculty members (79%) rated by all 224 third to fifth-year students (clerks) at outpatient departments of the AKU hospital over a one-year period. Repeated measures generalizability studies, correlations, concurrent validity of SETOC scores with best teacher nominations. Inter-rater G-coefficient and internal consistency of SETOC student ratings were 0.92 and 0.98. Average inter-item and inter-subscale correlations were 0.79 and 0.86. Comparing SETOC scores against "Best Teacher" nominations, sensitivity, specificity, positive and negative predictive values were greater than 0.84. Student ratings ranged from unsatisfactory (fourteen instructors) to outstanding (four instructors). Mean-scores for Learning-Milieu, Clinical-Teaching and General-Teaching were lower than those for Clinical-Competence and Global-Rating (p=0.000 for all). The SETOC elicited reliable and valid student ratings that can provide specific feedback to individual faculty with weak or outstanding teaching skills, and identify overall group shortcomings for faculty development.


Targeting MET and AXL overcomes resistance to sunitinib therapy in renal cell carcinoma.
Antiangiogenic therapy resistance occurs frequently in patients with metastatic renal cell carcinoma (RCC). The purpose of this study was to understand the mechanism of resistance to sunitinib, an antiangiogenic small molecule, and to exploit this mechanism therapeutically. We hypothesized that sunitinib-induced upregulation of the prometastatic MET and AXL receptors is associated with resistance to sunitinib and with more aggressive tumor behavior. In the present study, tissue microarrays containing sunitinib-treated and untreated RCC tissues were stained with MET and AXL antibodies. The low malignant RCC cell line 786-O was chronically treated with sunitinib and assayed for AXL, MET, epithelial-mesenchymal transition (EMT) protein expression and activation. Co-culture experiments were used to examine the effect of sunitinib pretreatment on endothelial cell growth. The effects of AXL and MET were evaluated in various cell-based models by short hairpin RNA or inhibition by cabozantinib, the multi-tyrosine kinases inhibitor that targets vascular endothelial growth factor receptor, MET and AXL. Xenograft mouse models tested the ability of cabozantinib to rescue sunitinib resistance. We demonstrated that increased AXL and MET expression was associated with inferior clinical outcome in patients. Chronic sunitinib treatment of RCC cell lines activated both AXL and MET, induced EMT-associated gene expression changes, including upregulation of Snail and β-catenin, and increased cell migration and invasion. Pretreatment with sunitinib enhanced angiogenesis in 786-0/human umbilical vein endothelial cell co-culture models. The suppression of AXL or MET expression and the inhibition of AXL and MET activation using cabozantinib both impaired chronic sunitinib treatment-induced prometastatic behavior in cell culture and rescued acquired resistance to sunitinib in xenograft models. In summary, chronic sunitinib treatment induces the activation of AXL and MET signaling and promotes prometastatic behavior and angiogenesis. The inhibition of AXL and MET activity may overcome resistance induced by prolonged sunitinib therapy in metastatic RCC.


Acute Physiological Responses to Strongman Training Compared to Traditional Strength Training.
Strongman training (ST) has become an increasingly popular modality, but data on physiological responses are limited. This study sought to determine physiological responses to an ST session compared to a traditional strength exercise training (RST) session. Ten healthy men (23.6 ± 27.5 years, 85.8 ± 10.3 kg) volunteered in a crossover design, where all participants performed an ST session, an RST session, and a resting session within 7 days apart. The ST consisted of sled drag, farmer's walk, 1 arm dumbbell clean and press, and tire flip at loads eliciting approximately 30 seconds of near maximal effort per set. The RST consisted of squat, deadlift, bench press, and power clean, progressing to 75% of 1 repetition maximum. Sessions were equated for approximate total set duration. Blood lactate and salivary testosterone were recorded immediately before and after training sessions. Heart rate, caloric expenditure, and substrate utilization were measured throughout the resting session, both training protocols and for 80 minutes after training sessions. Analyses were conducted to determine differences in physiological responses within and between protocols. No significant changes in testosterone occurred at any time point for either session. Lactate increased significantly immediately after both sessions. Heart rate, caloric expenditure, and substrate utilization were all elevated significantly during ST and RST. Heart rate and fat expenditure were significantly elevated compared to resting in both sessions' recovery periods; calorie and carbohydrate expenditures were not. Compared to RST, ST represents an equivalent physiological stimulus on key parameters indicative of potential training-induced adaptive responses. Such adaptations could conceivably include cardiovascular conditioning.


Everyday sovereignty: International experts, brokers and local ownership in peacebuilding Liberia.
The present article investigates how sovereignty is performed, enacted and constructed in an everyday setting. Based on fieldwork and interviews with international embedded experts about the elusive meaning of 'local ownership', we argue that while sovereignty may, indeed, be a model according to which the international community 'constructs' rogue or failed polities in 'faraway' places, this view overlooks that these places are still spaces in which contestations over spheres of authority take place every day, and thus also spaces in which sovereignty is constructed and reconstructed on a daily basis. Local ownership, then, becomes our starting point for tracing the processes of the everyday enactment of sovereignty. We make the case that sovereignty should not be reified, but instead be studied in its quotidian and dynamic production, involving the multiplicity of actors reflecting the active production of the state beyond its presumptive existence as a homogeneously organized, institutionalized and largely centralized bureaucracy.


Enhanced transepithelial antigen transport in intestine of allergic mice is mediated by IgE/CD23 and regulated by interleukin-4.
We previously described a system for enhanced transepithelial transport of antigen in which both the amount of specific antigen and its rate of transport were dramatically increased in intestine of sensitized rats compared with controls. This study investigated the essential components mediating antigen uptake in mice genetically deficient for interleukin (IL)-4 or CD23. Mice were actively or passively sensitized to horseradish peroxidase (HRP). Jejunal segments from control or sensitized mice were mounted in Ussing chambers and challenged with HRP from the luminal side. Tissues were processed for electron microscopy, and photomicrographs were analyzed for antigen uptake (location and area of HRP-containing endosomes). Immunohistochemistry and reverse-transcription polymerase chain reaction were used to detect epithelial CD23 expression. Actively sensitized IL-4(+/+), but not IL-4(-/-) mice, displayed increased transepithelial antigen transport and CD23 expression on enterocytes. Passively sensitized IL-4(+/+) and IL-4(-/-) mice displayed elevated antigen transport after transfer of immune serum but not if the serum was depleted of immunoglobulin (Ig) E or IL-4. IL-4 added to cultured IEC-4 cells up-regulated expression of CD23 messenger RNA. The augmented antigen uptake was inhibited by anti-CD23 and was absent in sensitized CD23(-/-) mice. Our studies indicate that IL-4 regulates IgE/CD23-mediated enhanced transepithelial antigen transport in sensitized mouse intestine.


Use of intestine-related biomarkers for detecting intestinal epithelial damage in neonatal calves with diarrhea.
To evaluate the usefulness of intestinal biomarkers in determining the presence of intestinal epithelial damage in neonatal calves with diarrhea caused by 4 etiologic agents. 40 neonatal calves that were healthy (n = 10) or had diarrhea (30). The study was a cross-sectional study. Results of hematologic analyses and serum concentrations of intestinal fatty acid-binding protein (I-FABP), liver fatty acid-binding protein (L-FABP), trefoil factor 3 (TFF-3), Claudin-3 (CLDN-3), γ-enteric smooth muscle actin (ACTG2), intestinal alkaline phosphatase (IAP), interleukin-8 (IL-8), platelet-activating factor (PAF), and leptin (LP) were compared among calves grouped according to whether they were healthy (control group; G-1) or had diarrhea caused by K99 Escherichia coli (G-2; n = 10), bovine rota- or coronavirus (G-3; 5 each), or Cryptosporidium spp (G-4; 10). Across the 3 time points at which blood samples were obtained and evaluated, the groups of calves with diarrhea generally had markedly higher mean serum concentrations of L-FABP, TFF-3, IAP, IL-8, and LP, compared with the control group. In addition, G-2 also consistently had markedly higher mean serum concentrations of I-FAB and ACTG2 and lower mean serum concentrations of CLDN-3, compared with the control group. Results indicated that degree of intestinal epithelial damage differed among calves grouped by the etiologic agent of diarrhea and that such damage might have been more severe in calves with diarrhea caused by K99 E coli. Additionally, our results indicated that serum concentrations of I-FABP, L-FABP, TFF-3, IAP, IL-8, ACTG2, LP, and CLDN-3 were useful biomarkers of intestinal epithelial damage in calves of the present study.


Identification and genomic characterization of influenza viruses with different origin in Mexican pigs.
Swine influenza is a worldwide disease, which causes damage to the respiratory system of pigs. The H1N1 and H3N2 subtypes circulate mainly in the swine population of Mexico. There is evidence that new subtypes of influenza virus have evolved genetically and have been rearranged with human viruses and from other species; therefore, the aim of our study was to identify and characterize the genetic changes that have been generated in the different subtypes of the swine influenza virus in Mexican pigs. By sequencing and analyzing phylogenetically the eight segments that form the virus genome, the following subtypes were identified: H1N1, H3N2, H1N2 and H5N2; of which, a H1N1 subtype had a high genetic relationship with the human influenza virus. In addition, a H1N2 subtype related to the porcine H1N2 virus reported in the United States was identified, as well as, two other viruses of avian origin from the H5N2 subtype. Particularly for the H5N2 subtype, this is the first time that its presence has been reported in Mexican pigs. The analysis of these sequences demonstrates that in the swine population of Mexico, circulate viruses that have suffered punctual-specific mutations and rearrangements of their proteins with different subtypes, which have successfully adapted to the Mexican swine population.


A simple synthetic route to polyoxovanadate-based organic-inorganic hybrids using EEDQ as an ester coupling agent.
A reaction strategy for the post-functionalization of hexavanadate derivatives is presented herein. In this study, five polyoxovanadate-based organic-inorganic hybrids TBA2[V6O13{(OCH2)3CCOO(CH2)15CH3}2] (2), TBA2[V6O13{(OCH2)3CCOO(CH2)8CH[double bond, length as m-dash]CH2}2] (3), TBA2[V6O13{(OCH2)3CCOOCH2CF3}2] (4), TBA2[V6O13{(OCH2)3CCOO(CH2CH2O)3CH3}2] (5), and TBA2[V6O13{(OCH2)3CCOO(CH2)12OH}2]·2CH3CH2OH (6) were successfully synthesized via esterification of carboxylic acid groups-containing TBA2[V6O13{(OCH2)3CCOOH}2] (1) with five alcohols possessing different functional groups. These hybrids were characterized by single crystal X-ray diffraction, IR, ESI-MS, 1H and 13C-NMR spectroscopies, and elemental analysis. Moreover, the formation of large vesicles was observed in a mixed solution of compound 2 due to its surfactant-like structure.


Temporal structure of first-year courses and success at course exams: comparison of traditional continual and block delivery of anatomy and chemistry courses.
To evaluate students' academic success at delivered in a traditional continual course, spread over the two semesters, or in alternating course blocks. We analyzed the data on exam grades for Anatomy and Chemistry courses in the first year of the curriculum for academic year 2001/02, with the traditional continual delivery of the courses (n=253 for chemistry and n=243 for anatomy), and academic year 2003/04, with block delivery of the courses (n=255 for Chemistry and n=260 for Anatomy). Grades from the final examination were analyzed only for students who sat the exam at the first available exam term and passed the course. For the Anatomy block course, grades at 2 interim written tests and 2 parts of the final exam (practical stage exam and oral exam) in each block were analyzed for students who passed all interim tests and the final exam. There were no differences between two types of course delivery in the number of students passing the final examination at first attempt. There was a decrease in passing percentage for the two Anatomy block course student groups in 2003/04 (56% passing students in block 1 vs 40% in block 2, P=0.014). There was an increase in the average grades from 2001/02 to 2003/04 academic year due to an increase in Chemistry grades (F1,399=18.4, P<0.001, 2 x 2 ANOVA). There was no effect of the sequence of their delivery (F1,206=1.8, P=0.182, 2 x 2 ANOVA). There was also a significant difference in grades on interim assessments of Anatomy when it was delivered in the block format (F3,85=28.8, P<0.001, between-within subjects 2 x 4 ANOVA). The type of course delivery was not associated with significant differences in student academic success in Anatomy and Chemistry courses in the medical curriculum. Students can successfully pass these courses when they are delivered either in a continual, whole year format or in a condensed time format of a course block, regardless of the number and type of courses preceding the block course.


Medial Gamma Knife thalamotomy for intractable pain.
OBJECTIVEAblative procedures are still useful in the treatment of intractable pain despite the proliferation of neuromodulation techniques. In the paper the authors present the results of Gamma Knife thalamotomy (GKT) in various pain syndromes.METHODSBetween 1996 and 2016, unilateral GKT was performed in 30 patients suffering from various severe pain syndromes in whom conservative treatment had failed. There were 20 women and 10 men in the study population, with a median age of 80 years (range 53-89 years). The pain syndromes consisted of 8 patients with classic treatment-resistant trigeminal neuralgia (TN), 6 with postherpetic TN, 5 with TN and constant pain, 1 with TN related to multiple sclerosis, 3 with trigeminal neuropathic pain, 4 with thalamic pain, 1 with phantom pain, 1 with causalgic pain, and 1 with facial pain. The median follow-up period was 24 months (range 12-180 months). Invasive procedures for pain release preceded GKT in 20 patients (microvascular decompression, glycerol rhizotomy, balloon microcompression, Gamma Knife irradiation of the trigeminal root, and radiofrequency thermolesion). The Leksell stereotactic frame, GammaPlan software, and T1- and T2-weighted sequences acquired at 1.5 T were used for localization of the targeted medial thalamus, namely the centromedian (CM) and parafascicularis (Pf) nucleus. The CM/Pf complex was localized 4-6 mm lateral to the wall of the third ventricle, 8 mm posterior to the midpoint, and 2-3 mm superior to the intercommissural line. GKT was performed using the Leksell Gamma Knife with an applied dose ranging from 145 to 150 Gy, with a single shot, 4-mm collimator. Pain relief after radiation treatment was evaluated. Decreased pain intensity to less than 50% of the previous level was considered successful.RESULTSInitial successful results were achieved in 13 (43.3%) of the patients, with complete pain relief in 1 of these patients. Relief was achieved after a median latency of 3 months (range 2-12 months). Pain recurred in 4 (31%) of 13 patients after a median latent interval of 24 months (range 22-30 months). No neurological deficits were observed.CONCLUSIONSThese results suggest that GKT in patients suffering from severe pain syndromes is a relatively successful and safe method that can be used even in severely affected patients. The only risk of GT for the patients in this study was failure of treatment, as no clinical side effects were observed.


Classification models for anticancer activity.
Despite significant research in understanding of neoplastic diseases, the success rate for oncology drugs is relatively very low. A major challenge before the scientific community is to design new chemical entities that will be highly selective for cancer cells so as to minimize side effects. Classification models (CMs) models play a prominent role in prediction of the biological properties of newly designed compounds before their synthesis and prevent non-optimal use of resources. Though correlation models far outnumber classification models for development of various therapeutic agents but the significance of classification models for development of anti-cancer agents can not be underestimated. Various techniques employed for development of classification models for anti-cancer activity have been briefly reviewed. Moreover, successful use of some of these classification techniques for the development of models for anti-proliferative activity has been illustrated using a data set comprising of 53 analogues of N-Benzoylated phenoxazines and phenothiazines. Resulting classification models with high degree of accuracy can play a vital role in providing lead structures for the development of novel anti-proliferative agents for cancer chemotherapy.


Characterization of the cytotoxic properties of the benzimidazole fungicides, benomyl and carbendazim, in human tumour cell lines and primary cultures of patient tumour cells.
The benzimidazoles, benomyl and carbendazim, are fungicides suggested to target microtubules. Benomyl is metabolized to carbendazim, which has already been explored as an anticancer drug in phase 1 clinical trials. We further characterized the cytotoxic properties of benomyl and carbendazim in 12 human cell lines and in primary cultures of patient tumour cells with the overall aims of elucidating mechanisms of action and anticancer activity spectrum. Cytotoxicity was assessed in the short-term fluorometric microculture cytotoxicity assay and was correlated with the activity of other anticancer drugs and gene expression assessed by cDNA microarray analysis. Benomyl was generally more potent than its metabolite, carbendazim. Both showed high drug activity correlations with several established and experimental anticancer drugs, but modest association with established mechanisms of drug resistance. Furthermore, these benzimidazoles showed high correlations with genes considered relevant for the activity of several mechanistically different standard and experimental anticancer drugs, indicating multiple and broad mechanisms of action. In patient tumour samples, benomyl tended to be more active in haematological compared with solid tumour malignancies, whereas the opposite was observed for carbendazim. In conclusion, benomyl and carbendazim show interesting and diverse cytotoxic mechanisms of action and seem suitable as lead compounds for the development of new anticancer drugs.


Immune deficiencies in children: an overview.
Primary immune deficiencies (PIDs) are disorders of the immune system that result in increased susceptibility to infectious disease, autoimmunity and malignancy. They are challenging to paediatricians as they can present anytime from birth to adolescence with a wide variety of signs and symptoms. It is important to diagnose PIDs promptly, especially more severe forms to prevent significant morbidity and mortality. However, significant challenges exist in deciding which children to investigate and when. We aim to give a basic understanding of the human immune system, the different presentations in a child that should alert a paediatrician about the possibility of PID and the possible underlying diagnosis. Additionally, we have developed a framework for a stepwise approach to investigating these children.


Deaths due to absence of an affordable antitoxin for plant poisoning.
There is a severe shortage of affordable antivenoms and antitoxins in the developing world. An anti-digoxin antitoxin for oleander poisoning was introduced in Sri Lanka in July, 2001, but because of its cost, stocks ran out in July, 2002. We looked at the effect of its introduction and withdrawal on case fatality, and determined its cost-effectiveness. The antitoxin strikingly reduced the case fatality; its absence resulted in a three-fold rise in deaths. At the present price of US2650 dollars per course, every life saved cost 10209 dollars and every life year cost 248 dollars. Reduction of the antitoxin's price to 400 dollars would reduce costs to 1137 dollars per life gained; a further reduction to 103 dollars would save money for every life gained. Treatments for poisoning and envenoming should be included in the present campaign to increase availability of affordable treatments in the developing world.


Extracellular matrix hydrogel derived from decellularized tissues enables endodermal organoid culture.
Organoids have extensive therapeutic potential and are increasingly opening up new avenues within regenerative medicine. However, their clinical application is greatly limited by the lack of effective GMP-compliant systems for organoid expansion in culture. Here, we envisage that the use of extracellular matrix (ECM) hydrogels derived from decellularized tissues (DT) can provide an environment capable of directing cell growth. These gels possess the biochemical signature of tissue-specific ECM and have the potential for clinical translation. Gels from decellularized porcine small intestine (SI) mucosa/submucosa enable formation and growth of endoderm-derived human organoids, such as gastric, hepatic, pancreatic, and SI. ECM gels can be used as a tool for direct human organoid derivation, for cell growth with a stable transcriptomic signature, and for in vivo organoid delivery. The development of these ECM-derived hydrogels opens up the potential for human organoids to be used clinically.


Immune and inflammation responses to a 3-day period of intensified running versus cycling.
Functional overreaching has been linked to alterations in immunity and host pathogen defense, but little is known as to whether or not running and cycling evoke different responses. This study compared inflammation, muscle damage and soreness, and innate immune function responses to a 3-day period of intensified exercise in trained long distance runners (N=13, age 34.4±2.4year) and cyclists (N=22, age 36.6±1.7year, P=0.452). Upper respiratory tract infection (URTI) symptomatology was monitored for 12weeks using the Wisconsin Upper Respiratory Symptom Survey (WURSS), and subjects from both athletic groups came to the lab during week five and exercised 2.5h/day for 3days in a row at 70% VO2max. Blood samples were collected before and after the 3-day period of exercise, with recovery samples collected 1-, 14-, and 38h-post-exercise. Samples were analyzed for muscle damage [creatine kinase (CK), myoglobin (MYO)], inflammation (CRP, IL-6, IL-8, IL-10, MCP), and innate immunity [granulocyte and monocyte phagocytosis (GR-PHAG and MO-PHAG) and oxidative burst activity (GR-OBA and MO-OBA)]. Runners compared to cyclists experienced significantly more muscle damage (CK 133% and MYO 404% higher post-3days exercise), inflammation (CRP 87%, IL-6 256%, IL 8 61%, IL-10 32%, MCP 29%), and delayed onset of muscle soreness (DOMS, 87%). The 3-day period of exercise caused significant downturns in GR-PHAG, MO-PHAG, GR-OBA, MO-OBA by 14- and 38h-recovery, but the pattern of change did not differ between groups. No group differences were measured for 12-week URTI severity (18.3±5.6 and 16.6±4.0, P=0.803) and symptom scores (33.4±12.6 and 24.7±5.8, P=0.477). These data indicate that a 3-day period of functional overreaching results in substantially more muscle damage and soreness, and systemic inflammation in runners compared to cyclists, but without group differences for 12-week URTI symptomatology and post-exercise decrements in innate immune function.


A novel mechanism of retrovirus inactivation in human serum mediated by anti-alpha-galactosyl natural antibody.
Type C retroviruses endogenous to various nonprimate species can infect human cells in vitro, yet the transmission of these viruses to humans is restricted. This has been attributed to direct binding of the complement component C1q to the viral envelope protein p15E, which leads to classical pathway-mediated virolysis in human serum. Here we report a novel mechanism of complement-mediated type C retrovirus inactivation that is initiated by the binding of "natural antibody" [Ab] (anti-alpha-galactosyl Ab) to the carbohydrate epitope Gal alpha 1-3Gal beta 1-4GlcNAc-R expressed on the retroviral envelope. Complement-mediated inactivation of amphotropic retroviral particles was found to be restricted to human and other Old World primate sera, which parallels the presence of anti-alpha-galactosyl natural Ab. Blockade or depletion of anti-alpha-galactosyl Ab in human serum prevented inactivation of both amphotropic and ecotropic murine retroviruses. Similarly, retrovirus was not killed by New World primate serum except in the presence of exogenous anti-alpha-galactosyl Ab. Enzyme-linked immunosorbent assays revealed that the alpha-galactosyl epitope was expressed on the surface of amphotropic and ecotropic retroviruses, and Western blot analysis further localized this epitope to the retroviral envelope glycoprotein gp70. Finally, down-regulation of this epitope on the surface of murine retroviral particle producer cells rendered them, as well as the particles liberated from these cells, resistant to inactivation by human serum complement. Our data suggest that anti-alpha-galactosyl Ab may provide a barrier for the horizontal transmission of retrovirus from species that express the alpha-galactosyl epitope to humans and to other Old World primates. Further, these data provide a mechanism for the generation of complement-resistant retroviral vectors for in vivo gene therapy applications where exposure to human complement is unavoidable.


Student self-assessment in a paediatric objective structured clinical examination.
The objective structured clinical examination (OSCE) has been recognized not only as a useful assessment tool but also as a valuable method of promoting student learning. Student self-assessment is also seen as a means of helping students recognize their strengths and weaknesses, understand the relevance of core learning objectives and to take more responsibility for each stage of their work The authors sought to evaluate the accuracy of medical student self-assessment of their performance in the paediatric clerkship OSCE and thus obtain preliminary data for use in programme strengthening. A self-administered questionnaire was completed by successive groups of students immediately after the OSCE at the end of each clerkship rotation. Students assessed their performance at each station, using a performance rating scale. Performance data were summarized using descriptive and non-parametric tests. Basic statistical analysis of the Likert items was conducted by calculatingfrequencies, means and standard deviations. Regression analysis was used to correlate self-reported rating and actual performance in each station. A p value of < 0.05 was considered significant. Eighty-one students (92%) completed the questionnaire. Fifty-eight (72%) of the students achieved greater than minimum competence in their overall scores. Significant positive correlation (p < 0.05) between student self-rating and actual score was noted- among the following stations: technical skills, cardiovascular examination, assessment of dysmorphism, dermatology, communication and photographic interpretation stations. Students overestimated their performance in the gastrointestinal examination, radiological and arterial blood gas interpretation. Students underestimated their performance in the following: respiratory system, examination of the head, developmental and nutritional assessment. The findings highlight the perceived strengths and weaknesses in clinical competence and self-assessment skills and provide direction for programme training needs.


[Influenza vaccination in Germany. A population-based cross-sectional analysis of three seasons between 2002 and 2005].
Influenza represents a considerable worldwide health problem. Vaccination is the only preventive measure, capable of reducing mortality and morbidity of influenza in all age groups. The objective of this study was to assess the extent of influenza vaccination coverage in the seasons 2002/2003, 2003/2004 and 2004/2005 in Germany, to understand the driving forces and barriers to vaccination, and to determine vaccination intentions for the subsequent winter. The authors conducted a random-sampling, telephone-based household survey among noninstitutionalized individuals representative of the population aged > or =14. The surveys for the different seasons used the same questionnaire and were subsequently pooled. Four target groups were determined for analysis: (1) persons aged > or =60; (2) people working in the medical field; (3) persons suffering from chronic illness; and (4) a composite group made up of persons aged > or =60 or working in the medical field or suffering from a chronic illness. The overall sample consisted of 5,990 people. The influenza vaccination coverage rate in Germany increased from 22.3% in season 2002/2003 to 25.1% in season 2003/2004 and increased again to 26.5% in season 2004/2005. This increase is statistically significant (p = 0.007). The composite group coverage rate increased from 40.0% (2002/2003) to 42.2% (2004/2005). The driving forces and barriers to vaccination did not change over the years. The most frequent reasons for being vaccinated given by vaccinees were: influenza considered to be a serious illness which people wanted to avoid, having received advice from the family doctor or nurse to be vaccinated, and not wanting to infect family and friends. Reasons for not being vaccinated mentioned by people who have never been vaccinated were: thinking about it, however, not being vaccinated in the end, not expecting to catch influenza, and not having received a recommendation from the family doctor to be vaccinated. Major encouraging factors to influenza vaccination were: recommendation by the family doctor or nurse, more available information on the vaccine regarding efficacy and tolerance, and more information available about the disease. The vaccination coverage rate increased by 4.2% over the three seasons. The family doctor is the most important source of encouragement for people to be vaccinated against influenza. It seems that the public would be more likely to be vaccinated if they had more information on the efficacy and tolerance of the vaccine, as well as the disease. The authors therefore suggest that family doctors be better informed on influenza vaccine and the disease itself, so that they can actively inform their patients on these topics.


Differences in maternal lineages of New Zealand Black mice defined by restriction endonuclease analysis of mitochondrial DNA and by expression of maternally transmitted antigen.
Two substrains of New Zealand Black (NZB) mice have been compared with respect to expression of a maternally transmitted cell surface antigen, Mta, defined by cloned cytolytic T cells, and for restriction enzyme polymorphisms of mitochondrial DNA (mtDNA). These independent assays of maternal cytoplasmic inheritance provide strong evidence for genetic contamination of the NZB/BlPt substrain (NZB/Bl mice from Michael Potter's separate colony at the National Institutes of Health), in which the typical NZB immunologic abnormalities are at least partially ameliorated. The decisive data are the restriction enzyme maps of mtDNA for NZB/BlPt, which were identical with those of the common "old inbred" strains and quite different from those of NZB/BlN (NZB/Bl mice from the breeding facility at the National Institutes of Health). It is probable that the contamination of the NZB/BlPt substrain is related to phenotypic changes in their autoimmune state. More interestingly, the data are consistent with, although they do not prove, involvement of the mitochondrial genome in expression of a cell surface molecule.


On learning, representing, and generalizing a task in a humanoid robot.
We present a programming-by-demonstration framework for generically extracting the relevant features of a given task and for addressing the problem of generalizing the acquired knowledge to different contexts. We validate the architecture through a series of experiments, in which a human demonstrator teaches a humanoid robot simple manipulatory tasks. A probability-based estimation of the relevance is suggested by first projecting the motion data onto a generic latent space using principal component analysis. The resulting signals are encoded using a mixture of Gaussian/Bernoulli distributions (Gaussian mixture model/Bernoulli mixture model). This provides a measure of the spatio-temporal correlations across the different modalities collected from the robot, which can be used to determine a metric of the imitation performance. The trajectories are then generalized using Gaussian mixture regression. Finally, we analytically compute the trajectory which optimizes the imitation metric and use this to generalize the skill to different contexts.


Rubella seroprevalence among young women taipei county after routine measles, mumps and rubella immunization program.
To better target rubella immunization efforts by determining the age-specific rubella seroprevalence and the related factors for children in Taiwan after the 1986 national vaccine policy. A cross-sectional study was conducted for these girls aged 10 years (1994 cohorts), 13 years (1991 cohorts), 16 years (1988 cohorts), and 19 to approximately 22 years (1982-1985 cohorts), respectively, in 2004 in Taipei county. Participants were given a questionnaire detailing family income, education level, ethnicity of mother, number of siblings, and history of rubella infection. Possible predictors of rubella seropositivity and differences in seroprevalence were calculated by multivariate logistic regression analysis. A total of 826 cases were recruited. The geometric mean titer of rubella IgG antibody was 60.7 +/- 2.1 IU/ml. Rubella seropositive rates were 282/287 (98.3%) in children aged 10 years, 234/235 (99.6%) in aged 13 years, 179/185 (96.8%) in aged 16 years, and 110/119 (92. 4%) in aged 19-22 years. Older age, low maternal education level, and foreign mothers were correlated with seronegativity (p < 0.05). The MMR vaccine program has induced effectively the immunity against rubella. However, to eliminate congenital rubella syndrome, more effort should be taken for the vaccination of susceptible adolescents and young women.


Does childhood schooling affect old age memory or mental status? Using state schooling laws as natural experiments.
The association between schooling and old age cognitive outcomes such as memory disorders is well documented but, because of the threat of reverse causation, controversy persists over whether education affects old age cognition. Changes in state compulsory schooling laws (CSL) are treated as natural experiments (instruments) for estimating the effect of education on memory and mental status among the elderly. Changes in CSL predict changes in average years of schooling completed by children who are affected by the new laws. These educational differences are presumably independent of innate individual characteristics such as IQ. CSL-induced changes in education were used to obtain instrumental variable (IV) estimates of education's effect on memory (n = 10,694) and mental status (n = 9751) for white, non-Hispanic US-born Health and Retirement Survey participants born between 1900 and 1947 who did not attend college. After adjustment for sex, birth year, state of birth and state characteristics, IV estimates of education's effect on memory were large and statistically significant. IV estimates for mental status had very wide confidence intervals, so it was not possible to draw meaningful conclusions about the effect of education on this outcome. Increases in mandatory schooling lead to improvements in performance on memory tests many decades after school completion. These analyses condition on individual states, so differences in memory outcomes associated with CSL changes cannot be attributed to differences between states. Although unmeasured state characteristics that changed contemporaneously with CSL might account for these results, unobserved genetic variation is unlikely to do so.


Both common and specialty mushrooms inhibit adhesion molecule expression and in vitro binding of monocytes to human aortic endothelial cells in a pro-inflammatory environment.
Cardiovascular disease (CVD) is a leading cause of mortality in the United States as well as globally. Epidemiological studies show that regular fruit and vegetable consumption reduces CVD risk, in part, due to antioxidant activity and immunomodulation since oxidative stress and inflammation are features of atherogenesis. Accumulating evidence also shows that dietary fungi, viz., mushrooms, can protect against chronic disease by altering inflammatory environments such as those associated with CVD although most research has focused on specialty mushrooms. In this study, we tested the ability of both common and specialty mushrooms to inhibit cellular processes associated with CVD. Human aortic endothelial cells (HAEC) were incubated overnight with control media with dimethylsulfoxide (DMSO) vehicle (1% v/v) or containing DMSO extracts of whole dehydrated mushrooms (0.1 mg/mL), which included Agaricus bisporus (white button and crimini), Lentinula edodes (shiitake), Pleurotus ostreatus (oyster), and Grifola frondosa (maitake). Monolayers were subsequently washed and incubated with medium alone or containing the pro-inflammatory cytokine IL-1beta (5 ng/mL) for 6 h to upregulate pro-atherosclerotic adhesion molecules (AM). AM expression was assayed by ELISA and binding of U937 human monocytes pre-loaded with fluorescent dye was determined. White button mushrooms consistently reduced (p < 0.05) VCAM-1, ICAM-1, and E-selectin-1 expression, whereas other test mushrooms significantly modulated AM expression singly, collectively, or combinatorially. All mushrooms, however, significantly reduced binding of monocytes to both quiescent and cytokine-stimulated monolayers. These data provide evidence that dietary mushrooms can inhibit cellular processes such as adhesion molecule expression and ultimate binding of monocytes to the endothelium under pro-inflammatory conditions, which are associated with CVD. As a result, these findings support the notion that dietary mushrooms can be protective against CVD.


Paracrine effects of oocyte secreted factors and stem cell factor on porcine granulosa and theca cells in vitro.
Oocyte control of granulosa and theca cell function may be mediated by several growth factors via a local feedback loop(s) between these cell types. This study examined both the role of oocyte-secreted factors on granulosa and thecal cells, cultured independently and in co-culture, and the effect of stem cell factor (SCF); a granulosa cell derived peptide that appears to have multiple roles in follicle development. Granulosa and theca cells were isolated from 2-6 mm healthy follicles of mature porcine ovaries and cultured under serum-free conditions, supplemented with: 100 ng/ml LR3 IGF-1, 10 ng/ml insulin, 100 ng/ml testosterone, 0-10 ng/ml SCF, 1 ng/ml FSH (granulosa), 0.01 ng/ml LH (theca) or 1 ng/ml FSH and 0.01 ng/ml LH (co-culture) and with/without oocyte conditioned medium (OCM) or 5 oocytes. Cells were cultured in 96 well plates for 144 h, after which viable cell numbers were determined. Medium was replaced every 48 h and spent medium analysed for steroids. Oocyte secreted factors were shown to stimulate both granulosa cell proliferation (P < 0.001) and oestradiol production (P < 0.001) by granulosa cells throughout culture. In contrast, oocyte secreted factors suppressed granulosa cell progesterone production after both 48 and 144 hours (P < 0.001). Thecal cell numbers were increased by oocyte secreted factors (P = 0.02), together with a suppression in progesterone and androstenedione synthesis after 48 hours (P < 0.001) and after 144 hours (P = 0.02), respectively. Oocyte secreted factors also increased viable cell numbers (P < 0.001) in co-cultures together with suppression of progesterone (P < 0.001) and oestradiol (P < 0.001). In granulosa cell only cultures, SCF increased progesterone production in a dose dependent manner (P < 0.001), whereas progesterone synthesis by theca cells was reduced in a dose dependent manner (P = 0.002). Co-cultured cells demonstrated an increase in progesterone production with increasing SCF dose (P < 0.001) and an increase in oestradiol synthesis at the highest dose of SCF (100 ng/ml). In summary, these findings demonstrate the presence of a co-ordinated paracrine interaction between somatic cells and germ cells, whereby oocyte derived signals interact locally to mediate granulosa and theca cell function. SCF has a role in modulating this local interaction. In conclusion, the oocyte is an effective modulator of granulosa-theca interactions, one role being the inhibition of luteinization.


Functional remodeling of benign human prostatic tissues in vivo by spontaneously immortalized progenitor and intermediate cells.
Tissue remodeling or regeneration is believed to initiate from multipotent stem and progenitor cells. We report here the establishment of two spontaneously immortalized adult non-tumorigenic human prostate epithelial cell lines, NHPrE1 and BHPrE1. NHPrE1 (CD133(high)/CD44(high)/OCT4(high)/PTEN(high)) was characterized as a putative progenitor cell, and BHPrE1 (p63(high)/p53(high)/p21(WAF1)(high)/RB(high)) was characterized as a putative epithelial intermediate cell. Genomic analysis demonstrated an abnormal karyotype with genomic rearrangements including PTEN amplification in NHPrE1 and CTNNB1 (beta-catenin) amplification in BHPrE1 cells. Embedded three-dimensional culture of NHPrE1 showed greater branching than BHPrE1. A tissue recombination-xenografting model was utilized to compare remodeling of human prostatic tissues in vivo. A series of tissue recombinants, made by mixing different ratios of human prostatic epithelial cells and inductive rat urogenital sinus mesenchyme, were grafted to the renal capsule of severe combined immunodeficient mice. Both cell lines were able to regenerate benign secretory ductal-acinar architecture in vivo, containing intact basal and luminal epithelial layers confirmed by the expression of appropriate CK profiles. Prostate-specific antigen, 15-lipoxygenase-2, androgen receptor, and NKX3.1 proteins were appropriately expressed in the regenerated epithelia. Regeneration of benign prostatic glandular structures could be achieved using as few as 10 NHPrE1 cells, whereas 200,000 BHPrE1 cells were required to achieve prostatic architecture. This suggests a greater proportion of progenitor/stem cells in NHPrE1 than in BHPrE1. These cell lines provide important data on progenitor and intermediate cell phenotypes and represent significant new tools for the elucidation of molecular mechanisms of human prostatic regeneration, pathogenesis, and carcinogenesis.


Nitrite in nitric oxide biology: cause or consequence? A systems-based review.
All life requires nitrogen compounds. Nitrite is such a compound that is naturally occurring in nature and biology. Over the years, the pharmacological stance on nitrite has undergone a surprising metamorphosis, from a vilified substance that generates carcinogenic nitrosamines in the stomach to a life-saving drug that liberates a protective agent (nitric oxide or NO) during hypoxic events. Nitrite has been investigated as a vasodilator in mammals for over 125 years and is a known by-product of organic nitrate metabolism. There has been a recent rediscovery of some of the vasodilator actions of nitrite in physiology along with novel discoveries which render nitrite a fundamental molecule in biology. Until recently nitrite was thought to be an inert oxidative breakdown product of endogenous NO synthesis but the past few years have focused on the reduction of nitrite back to NO in the circulation as a possible mechanism for hypoxic vasodilatation. Nitrite has evolved into an endogenous signaling molecule and regulator of gene expression that may not only serve as a diagnostic marker but also find its role as a potential therapeutic agent of cardiovascular disease. These data therefore warrant a reevaluation on the fate and metabolism of nitrite in biological systems. This review serves to encompass the history and recent evolution of nitrite, the compartment-specific metabolism of nitrite and its role in plasma as a biomarker for disease, the role of nitrite as a potential regulator of NO homeostasis, and the future of nitrite-based research.


Data Integration and Mining for Synthetic Biology Design.
One aim of synthetic biologists is to create novel and predictable biological systems from simpler modular parts. This approach is currently hampered by a lack of well-defined and characterized parts and devices. However, there is a wealth of existing biological information, which can be used to identify and characterize biological parts, and their design constraints in the literature and numerous biological databases. However, this information is spread among these databases in many different formats. New computational approaches are required to make this information available in an integrated format that is more amenable to data mining. A tried and tested approach to this problem is to map disparate data sources into a single data set, with common syntax and semantics, to produce a data warehouse or knowledge base. Ontologies have been used extensively in the life sciences, providing this common syntax and semantics as a model for a given biological domain, in a fashion that is amenable to computational analysis and reasoning. Here, we present an ontology for applications in synthetic biology design, SyBiOnt, which facilitates the modeling of information about biological parts and their relationships. SyBiOnt was used to create the SyBiOntKB knowledge base, incorporating and building upon existing life sciences ontologies and standards. The reasoning capabilities of ontologies were then applied to automate the mining of biological parts from this knowledge base. We propose that this approach will be useful to speed up synthetic biology design and ultimately help facilitate the automation of the biological engineering life cycle.


Leadership sharing in maternity emergency teams: a retrospective cohort study in simulation.
Shared leadership is associated with improved team performance in many domains, but little is understood about how leadership is shared spontaneously in maternity emergency teams, and if it is associated with improved team performance. A video analysis study of multidisciplinary teams attending a maternity emergency management course was performed at a simulation centre colocated with a tertiary maternity hospital. Sixteen teams responding to a simulated postpartum haemorrhage were analysed between November 2016 and November 2017. Videos were transcribed, and utterances coded for leadership type using a coding system developed a priori. Distribution of leadership utterances between team members was calculated using the Gini coefficient. Teamwork was assessed using validated tools and clinical performance was assessed by time to perform a critical intervention and a checklist of required tasks. There was a significant sharing of leadership functions across the team despite the traditional recommendation for a singular leader, with the dominant leader only accounting for 58% of leadership utterances. There was no significant difference in Auckland Team Assessment Tool scores between high and low leadership sharing teams (5.02 vs 4.96, p=0.574). Time to critical intervention was shorter in low leadership sharing teams (193 s vs 312 s, p=0.018) but checklist completion did not differ significantly. Teams with better clinical performance had fewer leadership utterances beyond the dominant two leaders compared with poorer performing teams. Leadership is spontaneously shared in maternity emergency teams despite the recommendation for singular leadership. Spontaneous leadership emerging from multiple team members does not appear to be associated with the improvements in team performance seen in other domains.


[The use of halotherapy for the rehabilitation of patients with acute bronchitis and a protracted and recurrent course].
Halotherapy was used for rehabilitation in 25 patients with acute bronchitis of long-standing and recurrent types. The main therapeutic action was ensured by aerodispersed medium saturated with dry highly dispersed sodium chloride aerosol, the required mass concentration being maintained in the range of 1 to 5 mg/m3. Therapy efficacy was controlled through assessment of clinical, functional, immunological and microbiological findings. Metabolic activity values were taken into consideration as well. Positive dynamics of the function indices in the clinical picture resulted from elimination of pathogenic agents, control of slowly running inflammatory lesions and stimulation of some immune system factors. Favourable changes in metabolic activity were present: normalization of serotonin excretion, marked decrease of unbalance in lipid peroxidation-antioxidant system.


Experimental and theoretical investigations of the inelastic and reactive scattering dynamics of O(3p) + D2.
This paper presents a combined experimental and theoretical study of the dynamics of O((3)P) + D(2) collisions, with emphasis on a center-of-mass (c.m.) collision energy of 25 kcal mol(-1). The experiments were conducted with a crossed-molecular-beams apparatus, employing a laser detonation source to produce hyperthermal atomic oxygen and mass spectrometric detection to measure the product angular and time-of-flight distributions. The novel beam source, which enabled these experiments to be conducted, contributed unique challenges to the experiments and to the analysis, so the experimental methods and approach to the analysis are discussed in detail. Three different levels of theory were used: (1) quasiclassical trajectories (QCT), (2) time-independent quantum scattering calculations based on high-quality potential surfaces for the two lower-energy triplet states, and (3) trajectory-surface-hopping (TSH) studies that couple the triplet surfaces with the lowest singlet surface using a spin-orbit Hamiltonian derived from ab-initio calculations. The latter calculations explore the importance of intersystem crossing in the dynamics. Both experiment and theory show that inelastically scattered O atoms scatter almost exclusively in the forward direction, with little or no loss of translational energy. For the reaction, O((3)P) + D(2) --> OD + D, the experiment shows that, on average, approximately 50% of the available energy goes into product translation and that the OD product angular distributions are largely backward-peaked. These results may be interpreted in light of the QCT and TSH calculations, leading to the conclusion that the reaction occurs mainly on triplet potential energy surfaces with, at most, minor intersystem crossing to a singlet surface. Reaction on either of the two low-lying reactive triplet surfaces proceeds through a rebound mechanism in which the angular distributions are backward-peaked and the product OD is both vibrationally and rotationally excited. The quantum scattering results are in good agreement with QCT calculations, indicating that quantum effects are relatively small for this reaction at a collision energy of 25 kcal mol(-1).


pH and population density in the regulation of animal cell multiplication.
Sparse and dense cultures of chick embryo cells were affected differently by pH. The rates of cell multiplication and of thymidine-(3)H incorporation into DNA of dense cultures were increased as the pH was increased from 6.6 to 7.6. At pH higher than 7.6 the rate of multiplication decreased slightly in the dense cultures, but the rate of thymidine-(3)H incorporation continued to increase. The discrepancy was due in part to cell death and detachment at very high pH, and in part to a more rapid uptake of thymidine-(3)H at very high pH. Sparse cultures were much less sensitive to pH reduction and, when a suitably conditioned medium was used to minimize cell damage, very sparse cultures grew almost as well at pH 6.7 as at higher pH. The rates of cell multiplication and thymidine-(3)H incorporation at low pH decreased in the initially sparse cultures before they reached confluent cell densities. There was no microscope evidence of direct contact between plasma membranes of cells at these densities although the parallel orientation indicated that the cells were influencing locally each other's behavior. Even at much higher cell densities, electron microscopy revealed large intercellular gaps partly filled with a fragmentary electron-opaque material suspected to be glycoprotein. Wounding experiments showed that pH affected cell migration in a manner similar to its effects on cell multiplication. Low pH inhibited cell migration, but those cells which migrated into the denuded region multiplied as rapidly at low pH as at high pH. The effects of pH on growth were correlated with effects on the uptake of 2-deoxyglucose-(3)H. Dense populations of cells inhibited by low pH were stimulated to incorporate thymidine-(3)H by the addition of small amounts of diethylaminoethyl-dextran. Rous sarcoma cells at high cell density were less sensitive to pH than were normal cells at the same density, but were more sensitive than sparse normal cultures. The results suggest that cell growth is inhibited through the combined effects of both lowered pH and high cell density on cell surface permeability.


The health worker recruitment and deployment process in Kenya: an emergency hiring program.
Despite a pool of unemployed health staff available in Kenya, staffing levels at most facilities were only 50%, and maldistribution of staff left many people without access to antiretroviral therapy (ART). Because in the current system it takes one to two years to fill vacant positions, even when funding is available, an emergency approach was needed to fast-track the hiring and deployment process. A stakeholder group was formed to bring together leaders from several sectors to design and implement a fast-track hiring and deployment model that would mobilize 830 additional health workers. This model used the private sector to recruit and deploy new health workers and manage the payroll and employment contracts, with an agreement from the government to transfer these staff to the government payroll after three years. The recruitment process was shortened to less than three months. By providing job orientation and on-time pay checks, the program increased employee retention and satisfaction. Most of the active roadblocks to changes in the health workforce policies and systems are 'human' and not technical, stemming from a lack of leadership, a problem-solving mindset and the alignment of stakeholders from several sectors. It is essential to establish partnerships and foster commitment and collaboration to create needed change in human resource management (HRM). Strengthening appointment on merit is one of the most powerful, yet simplest ways in which the health sector and governments that seek to tackle the challenges of corruption and poor governance can improve their image and efficiency. The quality and integrity of the public health sector can be improved only through professionalizing HRM, reformulating and consolidating the currently fragmented HR functions, and bringing all the pieces together under the authority and influence of HR departments and units with expanded scopes. HR staff must be specialists with strategic HR functions and not generalists who are confined to playing a restricted and bureaucratic role.


Structural basis for tRNA decoding and aminoacylation sensing by T-box riboregulators.
T-box riboregulators are a class of cis-regulatory RNAs that govern the bacterial response to amino acid starvation by binding, decoding and reading the aminoacylation status of specific transfer RNAs. Here we provide a high-resolution crystal structure of a full-length T-box from Mycobacterium tuberculosis that explains tRNA decoding and aminoacylation sensing by this riboregulator. Overall, the T-box consists of decoding and aminoacylation sensing modules bridged by a rigid pseudoknot structure formed by the mid-region domains. Stem-I and the Stem-II S-turn assemble a claw-like decoding module, while the antiterminator, Stem-III, and the adjacent linker form a tightly interwoven aminoacylation sensing module. The uncharged tRNA is selectively recognized by an unexpected set of favorable contacts from the linker region in the aminoacylation sensing module. A complex structure with a charged tRNA mimic shows that the extra moiety dislodges the linker, which is indicative of the possible chain of events that lead to alternative base-pairing and altered expression output.


Observer reliability of arteriovenous malformations grading scales using current imaging modalities.
The aim of this study was to examine observer reliability of frequently used arteriovenous malformation (AVM) grading scales, including the 5-tier Spetzler-Martin scale, the 3-tier Spetzler-Ponce scale, and the Pollock-Flickinger radiosurgery-based scale, using current imaging modalities in a setting closely resembling routine clinical practice. Five experienced raters, including 1 vascular neurosurgeon, 2 neuroradiologists, and 2 senior neurosurgical residents independently reviewed 15 MRI studies, 15 CT angiograms, and 15 digital subtraction angiograms obtained at the time of initial diagnosis. Assessments of 5 scans of each imaging modality were repeated for measurement of intrarater reliability. Three months after the initial assessment, raters reassessed those scans where there was disagreement. In this second assessment, raters were asked to justify their rating with comments and illustrations. Generalized kappa (κ) analysis for multiple raters, Kendall's coefficient of concordance (W), and interclass correlation coefficient (ICC) were applied to determine interrater reliability. For intrarater reliability analysis, Cohen's kappa (κ), Kendall's correlation coefficient (tau-b), and ICC were used to assess repeat measurement agreement for each rater. Interrater reliability for the overall 5-tier Spetzler-Martin scale was fair to good (ICC = 0.69) to extremely strong (Kendall's W = 0.73) on initial assessment and improved on reassessment. Assessment of CT angiograms resulted in the highest agreement, followed by MRI and digital subtraction angiography. Agreement for the overall 3-tier Spetzler-Ponce grade was fair to good (ICC = 0.68) to strong (Kendall's W = 0.70) on initial assessment, improved on reassessment, and was comparable to agreement for the 5-tier Spetzler-Martin scale. Agreement for the overall Pollock-Flickinger radiosurgery-based grade was excellent (ICC = 0.89) to extremely strong (Kendall's W = 0.81). Intrarater reliability for the overall 5-tier Spetzler-Martin grade was excellent (ICC > 0.75) in 3 of the 5 raters and fair to good (ICC > 0.40) in the other 2 raters. The 5-tier Spetzler-Martin scale, the 3-tier Spetzler-Ponce scale, and the Pollock-Flickinger radiosurgery-based scale all showed a high level of agreement. The improved reliability on reassessment was explained by a training effect from the initial assessment and the requirement to defend the rating, which outlines a potential downside for grades determined as part of routine clinical practice to be used for scientific purposes.


Genetic and chemical characterization of a mutant that disrupts synthesis of the lipopolysaccharide core tetrasaccharide in Rhizobium leguminosarum.
A 2-kb region that complements the Tn5-derived lipopolysaccharide (LPS) rough mutant Rhizobium leguminosarum RU301 was sequenced. Two open reading frames (ORFs) were identified. The first ORF (lpcA) is homologous to a family of bacterial sugar transferases involved in LPS core tetrasaccharide biosynthesis. ORF2 (lpcB), in which Tn5 transposed, has no significant homology to any DNA in the GenBank-EMBL databases. Chemical characterization of LPS produced by strain RU301 demonstrated that the 3-deoxy-D-manno-2-octulosonic acid (Kdo) residue which normally attaches the core tetrasaccharide to the O chain was missing, suggesting that IpcB may encode a CMP-Kdo:LPS Kdo transferase.


Impact of the Charter of Quebec Values on psychological well-being of francophone university students.
This paper discusses results from a pilot study conducted in the spring of 2014 among young adults living in Montreal. The main objective of this study was to assess the relation between perception of the Charter of Quebec Values, 1 self-identification, perception of intercommunity relations, perceived discrimination, and psychological well-being in young students enrolled in undergraduate or graduate programs of a francophone university in Montreal. A total of 441 students (30.5% male, 69.5% female) took part in a web survey designed by the research team. The data analyses and results suggest that the debate around the Charter of Quebec values was associated with a shift from a predominantly positive perception of intercommunity relations to a predominantly negative one, particularly among women, immigrants, and those who self-identified as cultural or religious minorities. In addition, more than 30% of participants reported having experienced some form of ethnic or religious discrimination since the Charter was released (personally or as a witness). This was particularly the case among immigrants, as well as those who self-identified as bicultural or from cultural or religious minority groups. This study's results thus highlight the exacerbation of intercommunity tensions linked to the public debate around identity and intercommunity relations in Quebec.


Image-based model of the spectrin cytoskeleton for red blood cell simulation.
We simulate deformable red blood cells in the microcirculation using the immersed boundary method with a cytoskeletal model that incorporates structural details revealed by tomographic images. The elasticity of red blood cells is known to be supplied by both their lipid bilayer membranes, which resist bending and local changes in area, and their cytoskeletons, which resist in-plane shear. The cytoskeleton consists of spectrin tetramers that are tethered to the lipid bilayer by ankyrin and by actin-based junctional complexes. We model the cytoskeleton as a random geometric graph, with nodes corresponding to junctional complexes and with edges corresponding to spectrin tetramers such that the edge lengths are given by the end-to-end distances between nodes. The statistical properties of this graph are based on distributions gathered from three-dimensional tomographic images of the cytoskeleton by a segmentation algorithm. We show that the elastic response of our model cytoskeleton, in which the spectrin polymers are treated as entropic springs, is in good agreement with the experimentally measured shear modulus. By simulating red blood cells in flow with the immersed boundary method, we compare this discrete cytoskeletal model to an existing continuum model and predict the extent to which dynamic spectrin network connectivity can protect against failure in the case of a red cell subjected to an applied strain. The methods presented here could form the basis of disease- and patient-specific computational studies of hereditary diseases affecting the red cell cytoskeleton.


Soft Drusen in Age-Related Macular Degeneration: Biology and Targeting Via the Oil Spill Strategies.
AMD is a major cause of legal blindness in older adults approachable through multidisciplinary research involving human tissues and patients. AMD is a vascular-metabolic-inflammatory disease, in which two sets of extracellular deposits, soft drusen/basal linear deposit (BLinD) and subretinal drusenoid deposit (SDD), confer risk for end-stages of atrophy and neovascularization. Understanding how deposits form can lead to insights for new preventions and therapy. The topographic correspondence of BLinD and SDD with cones and rods, respectively, suggest newly realized exchange pathways among outer retinal cells and across Bruch's membrane and the subretinal space, in service of highly evolved, eye-specific physiology. This review focuses on soft drusen/BLinD, summarizing evidence that a major ultrastructural component is large apolipoprotein B,E-containing, cholesterol-rich lipoproteins secreted by the retinal pigment epithelium (RPE) that offload unneeded lipids of dietary and outer segment origin to create an atherosclerosis-like progression in the subRPE-basal lamina space. Clinical observations and an RPE cell culture system combine to suggest that soft drusen/BLinD form when secretions of functional RPE back up in the subRPE-basal lamina space by impaired egress across aged Bruch's membrane-choriocapillary endothelium. The soft drusen lifecycle includes growth, anterior migration of RPE atop drusen, then collapse, and atrophy. Proof-of-concept studies in humans and animal models suggest that targeting the "Oil Spill in Bruch's membrane" offers promise of treating a process in early AMD that underlies progression to both end-stages. A companion article addresses the antecedents of soft drusen within the biology of the macula.


Expression profiling identifies genes involved in neoplastic transformation of serous ovarian cancer.
The malignant potential of serous ovarian tumors, the most common ovarian tumor subtype, varies from benign to low malignant potential (LMP) tumors to frankly invasive cancers. Given the uncertainty about the relationship between these different forms, we compared their patterns of gene expression. Expression profiling was carried out on samples of 7 benign, 7 LMP and 28 invasive (moderate and poorly differentiated) serous tumors and four whole normal ovaries using oligonucleotide microarrays representing over 21,000 genes. We identified 311 transcripts that distinguished invasive from benign tumors, and 20 transcripts that were significantly differentially expressed between invasive and LMP tumors at p < 0.01 (with multiple testing correction). Five genes that were differentially expressed between invasive and either benign or normal tissues were validated by real time PCR in an independent panel of 46 serous tumors (4 benign, 7 LMP, 35 invasive). Overexpression of SLPI and WNT7A and down-regulation of C6orf31, PDGFRA and GLTSCR2 were measured in invasive and LMP compared with benign and normal tissues. Over-expression of WNT7A in an ovarian cancer cell line led to increased migration and invasive capacity. These results highlight several genes that may play an important role across the spectrum of serous ovarian tumorigenesis.


Direct demonstration of instabilities in oxygen concentrations within the extravascular compartment of an experimental tumor.
To test the hypothesis that temporal variations in microvessel red cell flux cause unstable oxygen levels in tumor interstitium, extravascular oxygenation of R3230Ac mammary tumors grown in skin-fold window chambers was measured using recessed tip polarographic microelectrodes. Red cell fluxes in microvessels surrounding pO2 measurement locations were measured using fluorescently labeled red cells. Temporal pO2 instability was observed in all experiments. Median pO2 was inversely related to radial distance from microvessels. Transient fluctuations above and below 10 mm Hg were consistently seen, except in one experiment near the oxygen diffusion distance limit (140 microm) where pO2 fluctuations were <2 mm Hg and median pO2 was <5 mm Hg. Vascular stasis was not seen in these experiments. These results show that fluctuations in red cell flux, as opposed to vascular stasis, can cause temporal variations in pO2 that extend from perivascular regions to the maximum oxygen diffusion distance.


Micelles prepared from poly(N-isopropylacrylamide-co-tetraphenylethene acrylate)-b-poly[oligo(ethylene glycol) methacrylate] double hydrophilic block copolymer as hydrophilic drug carrier.
Self-assembled micelles obtained from a double hydrophilic block copolymer (DHBC) based on poly(N-isopropylacrylamide) (PNIPAAm) are a facile and green strategy compared to the conventional solvent exchange method. However, hydrophobic drug encapsulation in micelles from PNIPAAm-based DHBC has relatively low drug loading (DL) and encapsulation efficiency (EE). Intermolecular and intramolecular hydrogen bondings of PNIPAAm chains are formed to exclude hydrophobic drugs. Nevertheless, hydrogen bonding can be used for hydrophilic drug delivery. Therefore, we report micelles prepared from a poly(N-isopropylacrylamide-co-tetraphenylethene acrylate)-b-poly[oligo(ethylene glycol) methacrylate] [P(NIPAAm-co-TPE)-b-POEGMA] double hydrophilic block copolymer as a hydrophilic drug (thymopentin, TP5) carrier. The FTIR results confirm hydrogen bond formation between PNIPAAm chain and TP5. Micelles are obtained by simply increasing the temperature above the critical micelle temperature (CMT). The self-assembly behaviour of polymeric micelles is investigated by DLS, TEM and aggregation-induced emission (AIE) phenomenon. Cytotoxicity results indicate that the micelles are biocompatible. The in vitro prolonged drug release (from 6 min to several hours) and in vivo immunity enhancement indicate that the micelles formed by P(NIPAAm-co-TPE)-b-POEGMA DHBC are promising candidates as hydrophilic drug carriers, where hydrogen bonding is formed between PNIPAAm and drug.


Effects of skin surface temperature distribution of thermal manikin on clothing thermal insulation.
Effects of the distribution of skin surface temperature were investigated on thermal insulation of clothing for whole and each body part. The experiments were carried out with a thermal manikin in a climatic chamber. The two types of clothing ensemble were measured. The measurement with nude thermal manikin were also conducted. The three variations of skin temperature distribution were set with the thermal manikin. The values of the thermal insulation of nude skin surface (Ia), the total thermal insulation of clothing (It) and the basic thermal insulation of clothing (Icl) were measured with this thermal manikin under each skin temperature distribution. As a result, the values of Ia and It were not affected by skin temperature distributions of the range of typical experimental conditions. However, It is necessary to carefully use the values of the thermal insulation of clothing for the body parts, because these values were more influenced by the skin temperature distribution than those for the whole body.


Novel septoria speckled leaf blotch resistance loci in a barley doubled-haploid population.
The genetics of resistance to Septoria speckled leaf blotch (SSLB), caused by Septoria passerinii, was studied in the Leger × CIho9831 barley doubled-haploid population. The 140 lines in the population segregated as 102 resistant and 38 susceptible, approximating a 3:1 ratio. A recombination map was developed using diversity arrays technology and other molecular markers. Quantitative trait locus (QTL) analysis demonstrated that resistance is primarily conferred either by having the CIho9831 allele at a QTL on 6HS or by having the CIho9831 allele at both of two QTLs on 3H and 2HL. In addition, ≈1/16 of the lines were resistant for unidentified reasons. This model predicts a resistant/susceptible ratio of 11:5, which fits the phenotypic observations. Minor QTLs were detected on 2HS and 1H. DNA sequences of linked markers suggest that the 6HS, 3H, and 2HS QTLs are part of resistance gene clusters and that the 6HS and 3H QTLs share homology. The 6HS QTL is identical to or closely linked to the SSLB resistance locus Rsp4 and the 1H QTL to the Rsp2 or Rsp3 locus. The 3H and 2HS QTLs are unique and offer new opportunities for pyramiding resistance genes through marker-assisted breeding for resistance to S. passerinii.


Extra-oviductal expression of oviductal glycoprotein 1 in mouse: Detection in testis, epididymis and ovary.
Oviductal glycoprotein 1 (OVGP1), also called oviductin, is an oviduct-specific protein and is suggested to play a role in fertilization. Traditionally, Ovgp1 has been shown to be exclusively expressed by the oviduct; however, recent studies have demonstrated its expression in some cancers. This observation led us to hypothesize that Ovgp1 might have some extra-oviductal expression. In the current study, we evaluated the mRNA and protein expression of Ovgp1 in normal reproductive tissues of male and female mice. For the first time, we demonstrate that beyond the oviduct, Ovgp1 mRNA is expressed in the testis, epididymis and ovary, but not in the uterus, cervix, vagina, breast, seminal vesicles and prostate gland. In the testis, Ovgp1 mRNA was localized in the cells at the base of seminiferous tubules (most likely, Sertoli cells), while the protein was detected in the round and elongating spermatids. In the epididymis, Ovgp1 transcripts were localized in epididymal epithelium of the caput but not the corpus and cauda; OVGP1 protein was, however, not detected in any of the segments but was present in the epididymal sperm. In the ovary, Ovgp1 transcripts and protein were detected in the surface epithelium, granulosa cells of the preantral and the antral follicles and corpus luteum. In both, the ovary and oviduct, the expression of Ovgp1 was found to be higher at estrus stage than at diestrus stage. To the best of our knowledge, this is the first study demonstrating the extra-oviductal expression of Ovgp1. Our data suggests that, beyond fertilization, Ovgp1 might have specific roles in gonadal physiology. [Laheri S, Modi D and Bhatt P 2017 Extra-oviductal expression of oviductal.


Evaluation of the radiosensitivity of the oxygenated tumor cell fractions in quiescent cell populations within solid tumors.
Labeling of all proliferating cells in C57BL/6J mice bearing EL4 tumors was achieved by continuous administration of 5-bromo-2'-deoxyuridine (BrdU). Tumors were irradiated with γ rays at a high dose rate or a reduced dose rate at 1 h after the administration of pimonidazole. Assessment of the responses of quiescent and total ( = proliferating + quiescent) cell populations were based on the frequencies of micronucleation and apoptosis using immunofluorescence staining for BrdU. The response of the pimonidazole-unlabeled tumor cell fractions was assessed by means of apoptosis frequency using immunofluorescence staining for pimonidazole. The total cell fraction that was not labeled with pimonidazole showed significantly enhanced radiosensitivity. However, a significantly greater decrease in radiosensitivity, evaluated using a delayed assay or a decrease in radiation dose rate, was observed in the quiescent cell compared with the total cell population. Overall, the quiescent cell population showed significantly greater radioresistance and capacity to recover from radiation-induced damage than the total tumor cell population. Thus we believe that the subfraction of the quiescent tumor cell population that was not labeled with pimonidazole and that was probably oxygenated is a critical target in the control of solid tumors.


Real-time intraoperative ureter visualization with a novel Near-Infrared Ray Catheter during laparoscopic hysterectomy for gynecological cancer.
Ureteral injuries are well-known complications of gynecologic surgery, with a higher prevalence in laparoscopic surgery than in laparotomy [1]. The use of near-infrared fluorescent imaging navigation is currently being considered a novel method to identify the ureters intraoperatively and prevent ureteral injuries [2]. The Near-Infrared Ray Catheter (NIRC) fluorescent ureteral catheter is a newly developed device, containing a fluorescent resin that can be recognized by near-infrared irradiation. We found few reports on the use of this catheter in laparoscopic surgery for colon and rectal cancer [3, 4], but no reports in gynecologic surgery. We demonstrate the feasibility, safety, and potential usefulness of the real-time intraoperative visualization of the ureters using a novel NIRC fluorescent ureteral catheter in laparoscopic hysterectomy for endometrial cancer. A 30-year-old woman with early grade 1 endometrioid carcinoma was treated with medroxyprogesterone acetate for fertility preservation. After achieving complete response, she got pregnant and underwent cesarean section. The recurrence of atypical endometrial hyperplasia one year post-delivery prompted a total laparoscopic hysterectomy. Before the laparoscopic surgery began, the NIRC fluorescent ureteral catheters were placed in the ureters under the obtainment of informed consent from the patient. During the surgery, the catheters were successfully visualized by near-infrared fluorescence observation, which helped identify the ureters clearly and prevent ureteral injuries. This novel ureteral imaging navigation is expected to be an effective tool in cases of obesity, severe pelvic adhesion, deep infiltrating endometriosis, and malignancy in gynecologic laparoscopic surgery to clearly identify the ureter and to reduce the risk of ureteral injury.


Cross-Sectional Area of the Rotator Cuff Muscles in MRI - Is there Evidence for a Biomechanical Balanced Shoulder?
To provide in-vivo evidence for the common biomechanical concept of transverse and craniocaudal force couples in the shoulder that are yielded by both the rotator cuff muscles (RCM) and the deltoid and to quantitatively evaluate and correlate the cross-sectional areas (CSA) of the corresponding RCM as a surrogate marker for muscle strength using MRI. Fifty patients (mean age, 36 years; age range, 18-57 years; 41 male, 9 female) without rotator cuff tears were included in this retrospective study. Data were assessed by two readers. The CSA (mm2) of all rotator cuff muscles was measured on parasagittal T1-weighted FSE sequence at two different positions (at the established "y-position" and at a more medial slice in the presumably maximal CSA for each muscle, i.e., the "set position"). The CSA of the deltoid was measured on axial intermediate-weighted FSE sequences at three positions. CSA measurements were obtained using 1.5 Tesla MR-arthrographic shoulder. Pearson's correlation for the corresponding CSA of the force couple as well as was the intraclass correlation coefficient for the inter- and intra-reader agreement was calculated. The mean CSA was 770 mm2 (±167) and 841 mm2 (±191) for the supraspinatus (in the y- and set-positions, respectively) and 984 mm2 (±241) and 1568 mm2 (±338) for the infraspinatus. The mean CSA was 446 mm2 (±129) and 438 mm2 (±128) for the teres minor (in the y- and set-positions, respectively) and 1953 mm2 (±553) and 2343 mm2 (±587) for the subscapularis. The three measurements of the deltoid revealed a CSA of 3063 mm2 (±839) for the upper edge, 3829 mm2 (±836) for the lower edge and 4069 mm2 (±937) for the middle of the glenoid. At the set position Pearson's correlation of the transverse force couple (subscapularis/infraspinatus) showed a moderate positive correlation of r = 0.583 (p<0.0001) and a strong correlation when the CSA of the teres minor was added to the infraspinatus CSA (r = 0.665, p = 0.0008) and a strong positive correlation of the craniocaudal force couple (supraspinatus/deltoid) that ranged from r = 0.565-0.698 (p<0.0001). Inter-reader agreement (ranged from 0.841 to 0.997, p = 0.0007) and intra-reader agreement were excellent (ranged from 0.863 to 0.999, p = 0.0006). The significant correlation of the CSA of the RCM that form the transverse (subscapularis/infraspinatus-teres minor) and craniocaudal (supraspinatus/deltoid) force couple measured by MR-arthrography supports the biomechanical concept of a dynamically balanced shoulder in patients with an intact rotator cuff.


Heterogeneity of the Y chromosome in Afro-Brazilian populations.
Sixteen biallelic markers (SRY10831a, SRY10831b, SRY4064, SRY2627, 92R7, P2, P3, M34, M9, M3, M2, YAP, M60, M89, M213, M216) located in the nonrecombinant region of the Y chromosome were analyzed in 209 individuals belonging to six Brazilian populations: four Afro-Brazilian populations, one population of white European descendants, and one population of Japanese descendants. The results showed that most of the Y chromosomes of the Afro-Brazilians were from sub-Saharan Africa and that the proportion of Y chromosomes of European origin was greater than that of Y chromosomes of Amerindian origin. No typical African or Amerindian haplogroup was detected among Japanese individuals, and only one white individual showed a typical African haplogroup. Haplogroup P-92R7, which is highly frequent in the Portuguese and Italian populations, was the most frequent among whites (54%), and haplogroup K-M9, which shows wide geographic distribution and is absent in Africa, was the most frequent among Japanese individuals (65.6%). The two semi-isolated Afro-Brazilian populations showed the highest and the lowest genetic diversity, respectively. These differences probably reflect the effect of greater or smaller gene flow between a small isolated group and other populations. These findings show that the process of admixture does not occur homogeneously, with a tendency toward preferential marriages within the ethnic group and a clear direction in unions between European men and Amerindian or African women in the past. The results agree with historical and social data about the formation of the Brazilian population and reveal some of the factors that contribute to its heterogeneity.


Three-dimensional angle measurement based on propagation vector analysis of digital holography.
We propose what we believe to be a novel method for highly accurate three-dimensional (3D) angle measurement based on propagation vector analysis of digital holography. Three-dimensional rotations in space can be achieved by use of a CCD camera and a multifacet object, which reflects an incident wave into different directions. The propagation vectors of the reflected waves from the object can be extracted by analyzing the object spectrum of the recorded hologram. Any small rotation of the object will induce a change in the propagation vectors in space, which can then be used for 3D angle measurement. Experimental results are presented to verify the idea.


Cell plasticity in homeostasis and regeneration.
Over the past decades, genetic analyses performed in vertebrate and invertebrate organisms deciphered numerous cellular and molecular mechanisms deployed during sexual development and identified genetic circuitries largely shared among bilaterians. In contrast, the functional analysis of the mechanisms that support regenerative processes in species randomly scattered among the animal kingdom, were limited by the lack of genetic tools. Consequently, unifying principles explaining how stress and injury can lead to the reactivation of a complete developmental program with restoration of original shape and function remained beyond reach of understanding. Recent data on cell plasticity suggest that beside the classical developmental approach, the analysis of homeostasis and asexual reproduction in adult organisms provides novel entry points to dissect the regenerative potential of a given species, a given organ or a given tissue. As a clue, both tissue homeostasis and regeneration dynamics rely on the availability of stem cells and/or on the plasticity of differentiated cells to replenish the missing structure. The freshwater Hydra polyp provides us with a unique model system to study the intricate relationships between the mechanisms that regulate the maintenance of homeostasis, even in extreme conditions (starvation and overfeeding) and the reactivation of developmental programs after bisection or during budding. Interestingly head regeneration in Hydra can follow several routes according to the level of amputation, suggesting that indeed the homeostatic background dramatically influences the route taken to bridge injury and regeneration.


Role of src-like protooncogenes in lymphocyte proliferation.
Cross-linking of membrane-bound immunoglobulins, which are B cell antigen receptors, causes proliferation and differentiation of B cells or the inhibition of their growth. The receptor-mediated signalling involves tyrosine phosphorylation of cellular proteins. The Src-like protein-tyrosine kinase Lyn is expressed preferentially in B cells and is an intracytoplasmic constituent of the B cell antigen receptor complex. Cross-linking of membrane-bound immunoglobulin M with antibody induced rapid increases in the kinase activities of Lyn and Lyn-associated phosphatidylinositol-3 kinase. Cross-linking of B-cell antigen receptor also induced association of Lyn with an 85 kDa non-catalytic subunit of phosphatidylinositol-3 kinase. Thus, Lyn is functionally associated with membrane-bound immunoglobulin M, and seems to participate in B cell antigen receptor-mediated signalling. On the other hand, accumulating evidence shows that Fyn and Lck are involved in T-cell activation. Co-immunoprecipitation experiments showed that Fyn was physically associated with T cell antigen receptor-CD3 complex. Transient expression of actively mutated Fyn, having Phe-528 instead of Tyr-528, in Jurkat T cells stimulated the fos promoter and serum response element (SRE), suggesting that the Fyn kinase stimulates c-fos expression through SRE. An analogous set of experiments showed that introduction of the active Fyn alone had little effect on IL-2 promoter. However, it could stimulate transcription from this promoter when transfected cells were stimulated by a combination of Concanavalin A (ConA) and 12-O-tetradecanoylphorbol-13-acetate (TPA). Under the same conditions, normal Lck and Lyn could not stimulate IL-2 promoter.


Automatic quantification of juvenile zebrafish aggression.
Although aggression is a common symptom of psychiatric disorders the drugs available to treat it are non-specific and can have unwanted side effects. The zebrafish is an ideal model for aggression research. Zebrafish are small, amenable to genetic and pharmacological manipulation, and agonistic behaviour can be measured reliably. In this study we have established a novel setup to automatically quantify aggression and locomotion in one-month old juvenile zebrafish, a stage at which fish exhibit adult-like behaviour but are small so that one camera can film several animals. We have validated our novel software by comparison to manual quantification of behaviour, characterised the aggression of one-month old fish, and demonstrated that we can detect alterations to aggression caused by mutation or drug application. The ability to record up to 12 juvenile fish allows us to speed up and standardise data acquisition compared to studies of single fish. This setup appears to be suitable to screen for drugs that decrease zebrafish aggression as a first step toward developing novel treatments for this behaviour.


Differences in IgG Fc Glycosylation Are Associated with Outcome of Pediatric Meningococcal Sepsis.
Pediatric meningococcal sepsis often results in morbidity and/or death, especially in young children. Our understanding of the reasons why young children are more susceptible to both the meningococcal infection itself and a more fulminant course of the disease is limited. Immunoglobulin G (IgG) is involved in the adaptive immune response against meningococcal infections, and its effector functions are highly influenced by the glycan structure attached to the fragment crystallizable (Fc) region. It was hypothesized that IgG Fc glycosylation might be related to the susceptibility and severity of meningococcal sepsis. Because of this, the differences in IgG Fc glycosylation between 60 pediatric meningococcal sepsis patients admitted to the pediatric intensive care unit and 46 age-matched healthy controls were investigated, employing liquid chromatography with mass spectrometric detection of tryptic IgG glycopeptides. In addition, Fc glycosylation profiles were compared between patients with a severe outcome (death or the need for amputation) and a nonsevere outcome. Meningococcal sepsis patients under the age of 4 years showed lower IgG1 fucosylation and higher IgG1 bisection than age-matched healthy controls. This might be a direct effect of the disease; however, it can also be a reflection of previous immunologic challenges and/or a higher susceptibility of these children to develop meningococcal sepsis. Within the young patient group, levels of IgG1 hybrid-type glycans and IgG2/3 sialylation per galactose were associated with illness severity and severe outcome. Future studies in larger groups should explore whether IgG Fc glycosylation could be a reliable predictor for meningococcal sepsis outcome.IMPORTANCE Meningococcal sepsis causes significant mortality and morbidity worldwide, especially in young children. Identification of risk factors for a more fulminant infection would help to decide on appropriate treatment strategies for the individual patients. Immunoglobulin G (IgG) plays an essential role in humoral immune responses and is involved in the adaptive immune response against meningococcal infections. Of great influence on the receptor affinity of IgG is the N-glycan on its fragment crystallizable (Fc) portion. In the present study, we analyzed IgG glycosylation during the fast development of meningococcal sepsis in children, and we were able to identify glycosylation features that are different between meningococcal sepsis patients and healthy controls. These features might be indicative of a higher susceptibility to meningococcal sepsis. In addition, we found glycosylation features in the patients that were associated with illness severity and severe disease outcome, having the potential to serve as a disease outcome predictor.