[Quality of life in patients with temporomandibular disorders].
Oral disorders have a psycho-social impact on the quality of life, that can be measured with instruments as the Oral Health Impact Profile questionnaire (OHIP). Using the OHIP, we evaluated if and how the orofacial pain can affect the quality of life in temporomandibular disorders (TMD) patients. A transversal case-control study was carried out. Study subjects were patients referred to the Section of Prosthetic Dentistry and Temporomandibular Disorders of the University of Pavia (Italy). Subjects were recruited sequentially until the target of 124. The controls were 61 "pain free" subjects, who were recruited from the same clinic. In analyzing the data, the chi squared test was used for categorical data, and t test and one-way analysis of variance were used for numerical scores. The subjects in this study were predominantly females (83.9%). The mean age of subjects was 35.1 years (standard deviation= 14.0). The most frequently reported symptoms were pain in the temporomandibular joint (TMJ) (87.1%). The data showed that orofacial pain had an important impact on daily life (p<0.05) and that its most common outcomes were psychological. Comparison with a "pain free" population clearly indicated that orofacial pain and associated symptoms negatively affect the quality of life of TMD patients.


Sexual Reproduction in Aspergillus flavus Sclerotia: Acquisition of Novel Alleles from Soil Populations and Uniparental Mitochondrial Inheritance.
Aspergillus flavus colonizes agricultural commodities worldwide and contaminates them with carcinogenic aflatoxins. The high genetic diversity of A. flavus populations is largely due to sexual reproduction characterized by the formation of ascospore-bearing ascocarps embedded within sclerotia. A. flavus is heterothallic and laboratory crosses between strains of the opposite mating type produce progeny showing genetic recombination. Sclerotia formed in crops are dispersed onto the soil surface at harvest and are predominantly produced by single strains of one mating type. Less commonly, sclerotia may be fertilized during co-infection of crops with sexually compatible strains. In this study, laboratory and field experiments were performed to examine sexual reproduction in single-strain and fertilized sclerotia following exposure of sclerotia to natural fungal populations in soil. Female and male roles and mitochondrial inheritance in A. flavus were also examined through reciprocal crosses between sclerotia and conidia. Single-strain sclerotia produced ascospores on soil and progeny showed biparental inheritance that included novel alleles originating from fertilization by native soil strains. Sclerotia fertilized in the laboratory and applied to soil before ascocarp formation also produced ascospores with evidence of recombination in progeny, but only known parental alleles were detected. In reciprocal crosses, sclerotia and conidia from both strains functioned as female and male, respectively, indicating A. flavus is hermaphroditic, although the degree of fertility depended upon the parental sources of sclerotia and conidia. All progeny showed maternal inheritance of mitochondria from the sclerotia. Compared to A. flavus populations in crops, soil populations would provide a higher likelihood of exposure of sclerotia to sexually compatible strains and a more diverse source of genetic material for outcrossing.


Conservative approach in the treatment of renal trauma in children.
Our aim was to evaluate the role of a conservative approach in the treatment of renal trauma in the pediatric age group. A total of 31 pediatric patients with renal trauma (male = 25, female = 6) in whom we used a conservative approach in our clinic between the years 2002 and 2010 were included in the study. Patients were assessed according to the type of trauma, type of treatment, findings of radiological imaging, concomitant other organ injuries and complications occurring during follow-up. Surgery was required in just 4 of 31 pediatric patients who were treated with the conservative approach. Of the patients who underwent surgery, only one child needed nephrectomy. No complications were detected in the clinical and radiological follow-up of the patients who received the conservative approach. The main objective in the treatment of renal trauma occurring in childhood is to protect the kidneys. If the hemodynamic findings are stable, a conservative approach should be the first preferred method of treatment in every grade of renal trauma that can occur in childhood.


Open architecture time of flight 3D SWIR camera operating at 150 MHz modulation frequency.
In the past two decades 3-D cameras have proven to be one of the next revolutions in machine vision. However, these devices are still an emerging technology with a particularly narrow set of commercially available devices. In this paper, the concept and execution of the first short wavelength infrared (SWIR) time-of-flight (ToF) 3-D camera system operating at a wavelength of 1550 nm is presented. By decoupling the optical and electrical components of the system in an open architecture we not only surpass many of the limitations of an on-chip integrated solution, but also can easily change the imaging device based on the requirements of the application. We achieve modulation frequencies up to 150 MHz, which exceeds the conventional values currently published for other large format modulators by about five times. This increase in the modulation frequency allows for a TOF camera with significantly higher depth resolution, while the open architecture design allows for a highly reconfigurable device that can be modified for specific working conditions.


Developmental exposures of male rats to soy isoflavones impact Leydig cell differentiation.
Testicular Leydig cells, which are the predominant source of the male sex steroid hormone testosterone, express estrogen receptors (ESRs) and are subject to regulation by estrogen. Following ingestion, the two major isoflavones in soybeans, genistin and daidzin, are hydrolyzed by gut microflora to form genistein and daidzein, which have the capacity to bind ESRs and affect gene expression. Thus, the increasing use of soy-based products as nondairy sources of protein has raised concerns about the potential of these products to cause reproductive toxicity. In the present study, perinatal exposure of male rats to isoflavones induced proliferative activity in Leydig cells. Isoflavones have the capacity to act directly as mitogens in Leydig cells, because genistein treatment induced Leydig cell division in vitro. Genistein action regulating Leydig cell division involved ESRs, acting in concert with signaling molecules in the transduction pathway mediated by protein kinase B (AKT) and mitogen-activated protein kinase (MAPK). Enhanced proliferative activity in the prepubertal period increased Leydig cell numbers, which alleviated deficits in androgen biosynthesis and/or augmented serum and testicular testosterone concentrations in adulthood. Together, these observations indicate that the perinatal exposures of male rats to isoflavones affected Leydig cell differentiation, and they imply that including soy products in the diets of neonates has potential implications for testis function.


[Potential of the image analyzer "videotest" for performing microphotometric study in cytology].
Precision features of image analyser "Videotest" and possibilities of its applications for cytophotometric investigations have been analysed. It has been shown that a photometric feature is linear at least to optical density 1.1. Cytophotometry of the Feulgen stained rat hepatocyte nuclei has been performed. Coefficients of variation of DNA for 2c, 4c and 8c nuclei are 7.72, 7.65 and 6.50%, respectively, and ratios between average values of these nuclei are 1.00:2.06:4.12. Possible causes of errors of digital television equipment for cytophotometry are discussed.


Improved Production and Biophysical Analysis of Recombinant Silicatein-α.
Silicatein-α is a hydrolase found in siliceous sea sponges with a unique ability to condense and hydrolyse silicon-oxygen bonds. The enzyme is thus of interest from the perspective of its unusual enzymology, and for potential applications in the sustainable synthesis of siloxane-containing compounds. However, research into this enzyme has previously been hindered by the tendency of silicatein-α towards aggregation and insolubility. Herein, we report the development of an improved method for the production of a trigger factor-silicatein fusion protein by switching the previous hexahistidine tag for a Strep-II tag, resulting in 244-fold improvement in protein yield compared to previous methods. Light scattering and thermal denaturation analyses show that under the best storage conditions, although oligomerisation is never entirely abolished, these nanoscale aggregates of the Strep-tagged protein exhibit improved colloidal stability and solubility. Enzymatic assays show that the Strep-tagged protein retains catalytic competency, but exhibits lower activity compared to the His6-tagged protein. These results suggest that the hexahistidine tag is capable of non-specific catalysis through their imidazole side chains, highlighting the importance of careful consideration when selecting a purification tag. Overall, the Strep-tagged fusion protein reported here can be produced to a higher yield, exhibits greater stability, and allows the native catalytic properties of this protein to be assessed.


Therapeutic strategy in unresectable metastatic colorectal cancer.
While surgery is the cornerstone treatment for early-stage colorectal cancer, chemotherapy is the first treatment option for metastatic disease when tumor lesions are frequently not fully resectable at presentation. Mortality from colon cancer has decreased over the past 30 years, but there is still a huge heterogeneity in survival rates that can be mainly explained by patient and tumor characteristics, host response factors, and treatment modalities. The management of unresectable metastatic colorectal cancer is a global treatment strategy, which applies several lines of therapy, salvage surgery, maintenance, and treatment-free intervals. The individualization of cancer treatment is based on the evaluation of prognostic factors for survival (serum lactate dehydrogenase level, performance status), and predictive factors for treatment efficacy [Kirsten rat sarcoma viral oncogene homolog (KRAS) mutation status]. The available treatment modalities for metastatic colorectal cancer are chemotherapy (fluoropyrimidine, oxaliplatin, irinotecan), anti-angiogenic agents (e.g. bevacizumab), and anti-epidermal growth factor agents (cetuximab, panitumumab). The increasing number of active compounds dictates the strategy of trials evaluating these treatments either in combination or sequentially. Alternative outcomes that can be measured earlier than overall survival are needed to shorten the duration and reduce the size and cost of clinical trials.


Comparison of Listeria monocytogenes Exoproteomes from biofilm and planktonic state: Lmo2504, a protein associated with biofilms.
The food-borne pathogen Listeria monocytogenes is the causative agent of the severe human and animal disease listeriosis. The persistence of this bacterium in food processing environments is mainly attributed to its ability to form biofilms. The search for proteins associated with biofilm formation is an issue of great interest, with most studies targeting the whole bacterial proteome. Nevertheless, exoproteins constitute an important class of molecules participating in various physiological processes, such as cell signaling, pathogenesis, and matrix remodeling. The aim of this work was to quantify differences in protein abundance between exoproteomes from a biofilm and from the planktonic state. For this, two field strains previously evaluated to be good biofilm producers (3119 and J311) were used, and a procedure for the recovery of biofilm exoproteins was optimized. Proteins were resolved by two-dimensional difference gel electrophoresis and identified by electrospray ionization-tandem mass spectrometry. One of the proteins identified in higher abundance in the biofilm exoproteomes of both strains was the putative cell wall binding protein Lmo2504. A mutant strain with deletion of the gene for Lmo2504 was produced (3119Δlmo2504), and its biofilm-forming ability was compared to that of the wild type using the crystal violet and the ruthenium red assays as well as scanning electron microscopy. The results confirmed the involvement of Lmo2504 in biofilm formation, as strain 3119Δlmo2504 showed a significantly (P < 0.05) lower biofilm-forming ability than the wild type. The identification of additional exoproteins associated with biofilm formation may lead to new strategies for controlling this pathogen in food processing facilities.


The homeobox-containing gene Wariai regulates anterior-posterior patterning and cell-type homeostasis in Dictyostelium.
We have identified a Dictyostelium gene, Wariai (Wri), that encodes a protein with a homeobox and seven ankyrin repeats; both domains are required for function. A null mutation results in a more than doubling of the size of the prestalk O (pstO) compartment, one of the anterior prestalk compartments lying along the anterior-posterior axis of the migrating slug. There is a concomitant decrease in the more posterior prespore domain and no change in the more anterior prestalk A (pstA) and prestalk AB (pstAB) domains. wri null cells also have a morphological defect consistent with an increase in the pstO cell population. Wri itself is preferentially expressed in the pstA but not the pstO compartment, raising the possibility that Wri regulation of pstO compartment size is nonautonomous. Analysis of chimeric organisms is consistent with this model. Development in Dictyostelium is highly regulative, with cells within the prestalk and prespore populations being able to transdifferentiate into other cells to maintain proper cell-type proportioning. Our results suggest that Wri controls cell-type proportioning, possibly by functioning as a negative regulator of a pathway mediating pstO cell differentiation and controlling the mechanism of homeostasis regulating the size of one or more of the cell-type compartments. Our results also suggest that homeobox gene regulation of anterior-posterior axis patterning may have evolved prior to the evolution of metazoans.


The genomics of domestication special issue editorial.
Domestication has been of major interest to biologists for centuries, whether for creating new plants and animal types or more formally exploring the principles of evolution. Such studies have long used combinations of phenotypic and genetic evidence. Recently, the advent of a large number of genomes and genomic tools across a wide array of domesticated plant and animal species has reinvigorated the study of domestication. These genomic data, which can be easily generated for nearly any species, often provide great insight with or without a reference genome. The comparison of genome wide data from domestic and wild species has ignited a wave of insight into human, plant, and animal history with a new range of questions becoming accessible. With this in mind, this issue of Evolutionary Applications includes eleven papers covering a wide range of perspectives and methodologies relevant to understanding genomic variation under domestication.


Gas-liquid chromatographic determination of vitamin D in multivitamin preparations containing excess amounts of vitamin E.
Gas-liquid chromatographic (GLC) determination of vitamin D in multivitamin preparations containing excess amounts of vitamin E (a more than 2,500 weight ratio of dl-alpha-tocopheryl acetate to vitamin D) was investigated and a simplified routine method was established because the method reported previously (1) could not be applied to such special preparations. After applying the unsaponifiable matters of a sample to a phosphate-treated alumina column chromatography prepared according to MULDER et. al. (2), the eluate was evaporated and the residue was subjected to thin-layer chromatography (TLC) and GLC. When this method was applied to model preparations made by mixing vitamin D2 and dl-alpha-tocopheryl acetate (excess amounts), good results were obtained. Since the results on a commercial multivitamin preparation containing excess amounts of vitamin E were also satisfactory, it was confirmed that the proposed method could be used for simplified routine determinations.


A nontoxic system for 41.8 degrees C whole-body hyperthermia: results of a Phase I study using a radiant heat device.
A Phase I study of whole-body hyperthermia (WBH) (52 treatments/12 patients) was completed with no significant clinical toxicity. The study incorporated a thermal dose escalation scheme from 39.5 degrees-41.8 degrees C for up to 151 min. A radiant-heat device was utilized for producing WBH. During WBH, patients were sedated; endotracheal intubation was not required. No changes in cardiovascular, respiratory, hematological, or biochemical indices requiring clinical intervention occurred during the study. We conclude the radiant-heat device coupled with a defined pharmacological approach to WBH with appropriate patient screening yields a system for 41.8 degrees C WBH which is safe and efficient, is not labor intensive, and does not require general anesthesia and endotracheal intubation. This system is appropriate for a multimodality approach to various systemic cancers.


Patent ductus arteriosus: endovascular treatment in adult patient.
The patent ductus arteriosus (PDA) is a common type of congenital heart defect and its correction is simple when performed early in life. Surgery is performed using stitches or clips. In adults, the anomaly can lead to pulmonary hypertension and ventricular dysfunction. Surgery in adults is controversial and high-risk. This report describes an alternative endovascular approach in an adult patient.


dlk inhibits stem cell factor-induced colony formation of murine hematopoietic progenitors: Hes-1-independent effect.
Delta-like (dlk) is a family of transmembrane proteins containing epidermal growth factor-like repeat motifs homologous to the notch/delta/serrate family. Recent studies suggest that dlk is a negative regulator of adipocyte differentiation, a promoting factor of cobblestone area colony formation, and a molecule which influences stromal cell-pre-B cell interactions and augments cellularity of developing thymocytes. However, the role of dlk in regulating the growth and differentiation of hematopoietic progenitors remains unclear. In the present study, we examined the effect of dlk on the proliferation of murine hematopoietic progenitors by hematopoietic growth factors. Soluble dlk-IgG Fc chimeric protein completely inhibited the colony formation of lineage-marker negative (Lin-) bone marrow cells by GM-CSF, G-CSF, or macrophage-CSF (M-CSF) in the presence of stem cell factor (SCF). However, dlk failed to inhibit the colony formation of Lin- bone marrow cells by CSF, as described above, or M-CSF plus interleukin 3. Furthermore, dlk failed to inhibit the colony formation of Hes-1-null fetal liver cells by M-CSF in the presence of SCF. These findings suggest that dlk is an important regulator of hematopoietic progenitor proliferation. Depending on the presence of SCF, dlk may act as a growth inhibitor, although dlk signaling does not mediate Hes-1 transcription factor.


Chemotactic responses to an attractant and a repellent by the polar and lateral flagellar systems of Vibrio alginolyticus.
Chemotactic responses in Vibrio alginolyticus, which has lateral and polar flagellar systems in one cell, were investigated. A lateral-flagella-defective (Pof+ Laf-) mutant, which has only a polar flagellum, usually swam forward by the pushing action of its flagellum and occasionally changed direction by backward swimming. When the repellent phenol was added, Pof+ Laf- cells moved frequently forward and backward (tumbling state). The tumbling was derived from the frequent changing between counter-clockwise and clockwise (CW) rotation of the flagellar motor, as was confirmed by the tethered-cell method. Furthermore, we found that the tumbling cells did not adapt to the phenol stimulus. When the attractant serine was added, the phenol-treated cells ceased tumbling and swam smoothly, adapting to the attractant stimulus after several minutes. We isolated chemotaxis-defective (Che-) mutants from the Pof+ Laf- mutant; the tumbling mutants were not isolated. One interesting mutant swam backwards continuously, with its flagellum leading the cell and its flagellar motor rotating CW continuously. A polar-flagella-defective mutant (Pof- Laf+) stopped swimming after phenol addition and then recovered swimming ability within 10 min, indicating that lateral flagella can adapt to the repellent stimulus. This may represent a functional difference between the two flagellar systems in Vibrio cells, and between the chemotaxis systems affecting the two types of flagella.


Evolution and current status of the Whipple procedure: an update for gastroenterologists.
The Whipple procedure has undergone a remarkable gradual evolution in the last 20 years, of which many gastroenterologists are unaware. Improvements in staging, particularly staging laparoscopy with ultrasonography, have reduced the incidence of negative laparotomies. The forbidding mortality of pancreaticoduodenectomy, approximately 20% just a generation ago, has decreased precipitously in high-volume referral centers. Near zero mortality rates are now common. Morbidity and length of stay have also been reduced. Cardiac and pulmonary complications have been markedly reduced, whereas others such as pancreatic fistula still remain a problem. Modifications of the procedure have been introduced to improve long-term outcome of pancreatic cancer and to lessen digestive sequelae. Total pancreatectomy and large regional excisions did not improve results. However, 5-year survival rates of 20% are now reported by several centers for adenocarcinoma of the pancreas, and long-term survival rates for other periampullary tumors are approximately 40%. Pylorus-sparing procedures can be performed and may lessen postoperative sequelae. The clinical consequences of improved results are that large numbers of procedures are being performed at specialty centers, providing the opportunity to perform clinical trials, and that the procedure is used more widely, for instance, in benign diseases such as chronic pancreatitis.


Chemoselective Sequential Click Ligations Directed by Enhanced Reactivity of an Aromatic Ynamine.
Aromatic ynamines or N-alkynylheteroarenes are highly reactive alkyne components in Cu-catalyzed Huisgen [3 + 2] cycloaddition ("click") reactions. This enhanced reactivity enables the chemoselective formation of 1,4-triazoles using the representative aromatic ynamine N-ethynylbenzimidazole in the presence of a competing aliphatic alkyne substrate. The unique chemoselectivity profile of N-ethynylbenzimidazole is further demonstrated by the sequential click ligation of a series of highly functionalized azides using a heterobifunctional diyne, dispelling the need for alkyne protecting groups.


[Laboratory methods of controlling the effectiveness of antibacterial therapy].
The serum antibacterial activity (SAA) against causative agent isolated after the use of antibiotics was studied in 68 patients with pyoseptic diseases. The SAA ranged from 1:2 to 1:512 and depended on the antibiotic sensitivity of the causative agents. Antibiotic therapy was effective, when the SAA was equal to 1:8-1:512. With the use of monotherapy the adequacy of the regimens was controlled by the relation between the maximal blood level of the antibiotic and its MIC for the causative agent. The favourable clinical effect of the treatment with aminoglycosides and beta-lactams corresponded to the SAA exceeding 4.


Peak force visible microscopy.
The optical responses of molecules and materials provide a basis for chemical measurement and imaging. The optical diffraction limit in conventional light microscopy is exceeded by mechanically probing optical absorption through the photothermal effect with atomic force microscopy (AFM). However, the spatial resolution of AFM-based photothermal optical microscopy is still limited, and the sample surface is prone to damage from scratching due to tip contact, particularly for measurements on soft matter. In this article, we develop peak force visible (PF-vis) microscopy for the measurement of visible optical absorption of soft matter. The spatial resolution of PF-vis microscopy is demonstrated to be 3 nm on green fluorescent protein-labeled virus-like particles, and the imaging sensitivity may approach a single protein molecule. On organic photovoltaic polymers, the spatial distribution of the optical absorption probed by PF-vis microscopy is found to be dependent on the diffusion ranges of excitons in the donor domain. Through finite element modeling and data analysis, the exciton diffusion range of organic photovoltaics can be directly extracted from PF-vis images, saving the need for complex and delicate sample preparations. PF-vis microscopy will enable high-resolution nano-imaging based on light absorption of fluorophores and chromophores, as well as deciphering the correlation between the spatial distribution of photothermal signals and underlying photophysical parameters at the tens of nanometer scale.


Profiling of the Human Natural Killer Cell Receptor-Ligand Repertoire.
Natural killer (NK) cells are among the first responders to viral infections. The ability of NK cells to rapidly recognize and kill virally infected cells is regulated by their expression of germline-encoded inhibitory and activating receptors. The engagement of these receptors by their cognate ligands on target cells determines whether the intercellular interaction will result in NK cell killing. This protocol details the design and optimization of two complementary mass cytometry (CyTOF) panels. One panel was designed to phenotype NK cells based on receptor expression. The other panel was designed to interrogate expression of known ligands for NK cell receptors on several immune cell subsets. Together, these two panels allow for the profiling of the human NK cell receptor-ligand repertoire. Furthermore, this protocol also details the process by which we stain samples for CyTOF. This process has been optimized for improved reproducibility and standardization. An advantage of CyTOF is its ability to measure over 40 markers in each panel, with minimal signal overlap, allowing researchers to capture the breadth of the NK cell receptor-ligand repertoire. Palladium barcoding also reduces inter-sample variation, as well as consumption of reagents, making it easier to stain samples with each panel in parallel. Limitations of this protocol include the relatively low throughput of CyTOF and the inability to recover cells after analysis. These panels were designed for the analysis of clinical samples from patients suffering from acute and chronic viral infections, including dengue virus, human immunodeficiency virus (HIV), and influenza. However, they can be utilized in any setting to investigate the human NK cell receptor-ligand repertoire. Importantly, these methods can be applied broadly to the design and execution of future CyTOF panels.


A polysomal ribonuclease involved in the destabilization of albumin mRNA is a novel member of the peroxidase gene family.
We have purified an approximately 60 kDa endoribonuclease from Xenopus liver polysomes with properties expected for a messenger RNase involved in the estrogen-regulated destabilization of serum protein mRNAs (Dompenciel et al., 1995, J Biol Chem 270:6108-6118). The present report describes the cloning of this protein and its identification as a novel member of the peroxidase gene family. This novel enzyme, named polysomal RNase 1, or PMR-1 has 57% sequence identity with myeloperoxidase, and like that protein, appears to be processed from a larger precursor. Unlike myeloperoxidase, however, PMR-1 lacks N-linked oligosaccharide, heme, and peroxidase activity. Western blot and immunoprecipitation experiments using epitope-specific antibodies to the derived protein sequence confirm the identity of the cloned cDNA to the protein originally isolated from polysomes. The 80 kDa pre-PMR-1 expressed in a recombinant baculovirus was not processed to the 60 kDa form in Sf9 cells and lacks RNase activity. However, the baculovirus-expressed mature 60-kDa form of the enzyme has RNase activity. The recombinant protein is an endonuclease that shows selectivity for albumin versus ferritin mRNA. While it does not cleave at consensus APyrUGA elements, recombinant PMR-1 generates the same minor cleavage products from albumin mRNA as PMR-1 purified from liver. Finally, we show estrogen induces only a small increase in the amount of PMR-1. This result is consistent with earlier data suggesting estrogen activates mRNA decay through a posttranslational pathway.


Comparison of TTS-fentanyl with sustained-release oral morphine in the treatment of patients not using opioids for mild-to-moderate pain.
This randomised, multicentre, direct open comparative trial evaluated the efficacy, treatment convenience, tolerability and safety aspects of transdermal therapeutic system (TTS)-fentanyl and sustained-release oral morphine (SRM) in both opioid-naïve patients with moderate-to-severe cancer-related pain and in patients who had already been using opioids for mild-to-moderate pain. The two treatment groups were run in parallel. Special attention was paid to constipation, nausea/vomiting, drowsiness and respiratory depression. The 131 enrolled patients started the 4-week treatment at low doses of opioid (25 microg/h TTS-fentanyl for 3 days or 30 mg SRM every 12 h) and were individually titrated. Tolerability, efficacy and safety were assessed throughout the study period. Frequency of constipation was the primary study variable and accordingly the study was powered for this. Both patients and investigators made a global treatment evaluation. TTS-fentanyl and SRM were shown to be equally effective. Pain control and sleep quality improved with both treatments. None of the patients developed respiratory depression. Statistically significantly more patients in the SRM treatment group discontinued the trial prematurely (59% vs 27%; p < 0.001), particularly due to adverse events (36% vs 4%; p < 0.001). Fewer patients in the TTS-fentanyl than in the SRM treatment group reported constipation during the trial. This finding was statistically significant after 1 week of treatment (27% vs 57%; p = 0.003). The favourable tolerability profile of TTS-fentanyl was also reflected in both the patient and the investigator global evaluation of the treatment. Patient assessment favoured TTS-fentanyl treatment in terms of a significantly lower rate of troublesome side-effects ('quite a bit' to 'very much' troublesome side-effects in 14% vs 36% of patients; p = 0.003) and less interruption of daily activities (absence of any interruption of daily activities in 88% vs 63% of patients; p = 0.012). Investigators scored TTS-fentanyl as significantly better with respect to 'side-effects' (p = 0.039) and 'overall impression' (p = 0.013). Sub-analyses of opioid-naïve users gave similar results. These data indicate that TTS-fentanyl, when used as an opioid of first choice in the treatment of cancer-related pain, is as effective as, but better tolerated than, SRM, including in opioid-naïve patients.


Unique effect of arachidonic acid on human neutrophil TNF receptor expression: up-regulation involving protein kinase C, extracellular signal-regulated kinase, and phospholipase A2.
Arachidonic acid (AA) regulates the function of many cell types, including neutrophils. Although much emphasis has been placed on agonist-induced down-regulation of TNFR, our data show that AA caused a rapid (10-20 min) and dose-dependent (0.5-30 micro M) increase in the surface expression of both classes of TNFR (TNFR1 and TNFR2) on human neutrophils. This increased TNFR expression correlated with an increase in TNF-induced superoxide production. In contrast, the omega3 fatty acids eicosapentaenoic acid, docosahexaenoic acid, and linolenic acid failed to stimulate TNFR expression. Although fMLP and LPS reduced the neutrophil expression of TNFR, when pretreated with AA, fMLP caused an increase in TNFR expression. Consistent with this result was the finding that AA prevented the fMLP-induced receptor release in neutrophil cultures. AA also caused an increase in TNFR expression in matured HL-60 cells (neutrophil-like cells), but a decrease in nonmatured cells and HUVEC. The AA effects were independent of the lipoxygenase and cyclooxygenase pathways, but dependent on protein kinase C, the extracellular signal-regulated kinases 1 and 2, and cytosolic phospholipase A(2). The data demonstrate a unique effect of AA in the inflammatory reaction, through its action on neutrophil TNFR expression, and suggest that AA may regulate the response of neutrophils to TNF by altering its receptor number.


The Effects of 3 Weeks of Uphill and Downhill Walking on Blood Lipids and Glucose Metabolism in Pre-Diabetic Men: A Pilot Study.
The prevention of type 2 diabetes in persons at risk for diabetes is of utmost importance. Physical activity in general and even exercises at moderate intensities such as walking significantly reduce the risk of the development of type 2 diabetes. However, it is still a matter of debate whether lipids and glucose metabolism are differently affected by regular concentric (e.g., uphill walking) and eccentric (e.g., downhill walking) endurance exercise. The aim of this study was to investigate the effects of short-term (3 weeks) uphill and downhill walking on glucose metabolism and blood lipids in pre-diabetic middle-aged men in a real world setting. The study was designed as an investigator-initiated 2 group random selection pre-test post-test trial. Sixteen pre-diabetic men (age: 56.9 ± 5.1 years; BMI: 28.1 ± 2.3 kg·m-2) performed 9 uphill (n = 8) or 9 downhill (n = 8) walking sessions within 3 weeks. The primary outcomes were the markers of glucose metabolism and blood lipids measured before and after the training period. After uphill walking glucose tolerance (area under the curve of the oral glucose tolerance test: -43.25 ± 53.12 mg·dl-1; p = 0.05; effect size: 0.81), triglycerides (-48.75 ± 54.49 mg·dl-1; p = 0.036; effect size: 0.89), HDL-C (+7.86 ± 9.54 mg·dl-1; p = 0.05; effect size: 0.82) and total cholesterol/HDL-C ratio (-0.58 ± 0.41; p = 0.012; effect size: 1.39) had significantly improved. No significant metabolic adaptations were found after downhill walking. However, when adjusted for estimated energy expenditure, uphill and downhill walking had equal effects on almost all metabolic parameters. Moreover, the magnitude of the baseline impairments of glucose tolerance was significantly related to the extent of change in both groups. Depending on the fitness level and individual preferences both types of exercise may be useful for the prevention of type 2 diabetes and disorders in lipid metabolism.


Losing Weight on Reality TV: A Content Analysis of the Weight Loss Behaviors and Practices Portrayed on The Biggest Loser.
A number of weight loss-related reality television programs chronicle the weight loss experience of obese individuals in a competitive context. Although highly popular, such shows may misrepresent the behavior change necessary to achieve substantial weight loss. A systematic, quantitative content analysis of Seasons 10-13 (n = 66 episodes) of The Biggest Loser was conducted to determine the amount of time and number of instances that diet, physical activity, or other weight management strategies were presented. The average episode was 78.8 ± 15.7 min in length. Approximately 33.3% of an episode, representing 1,121 segments, portrayed behavioral weight management-related content. Within the episode time devoted to weight management content, 85.2% was related to physical activity, 13.5% to diet, and 1.2% to other. Recent seasons of The Biggest Loser suggest that substantial weight loss is achieved primarily through physical activity, with little emphasis on modifying diet and eating behavior. Although physical activity can impart substantial metabolic health benefits, it may be difficult to create enough of an energy deficit to induce significant weight loss in the real world. Future studies should examine the weight loss attitudes and behaviors of obese individuals and health professionals after exposure to reality television shows focused on weight loss.


About collagen, a tribute to Yves Bouligand.
Yves Bouligand's analysis of the organizations of biological materials in relation to those of liquid crystals enabled the development of the idea that physical forces exerting their actions under strong spatial constraints determine the structures and morphologies of these materials. The different levels of organization in collagen have preoccupied him for a long time. We present here our recent works in this domain that we were still discussing with him a few months before his death at the age of 76 on 21 January 2011. After recalling the hierarchical set of structures built by collagen molecules, we analyse them, exploiting the properties of the curved space of the hypersphere and of the algorithm of phyllotaxis. Those two geometrical concepts can be proposed as structural archetypes founding the polymorphism of this complex material of biological origin.


Optical imaging of breast cancer using hemodynamic changes induced by valsalva maneuver.
To investigate whether changes in hemodynamics induced by Valsalva maneuver can be exploited for detecting and characterizing breast lesions by optical mammography. 30 women underwent optical imaging of the breast using a DYNOT 232 system and performing Valsalva maneuvers prior to biopsy. Changes in light absorption due to changes in oxyhemoglobin and deoxyhemoglobin concentrations were recorded volumetrically and in a time-resolved manner. The parameters full width at half maximum (FWHM), time to ten (TTT), and peak amplitude (PA) of the reconstructed concentration time curves yielded color-coded maps of the breast which were separately evaluated by two experienced readers for detection rate, degree of visibility, and detection of additional lesions. ROC analysis was performed with the evaluation results. 10 patients were excluded from analysis due to artifacts or inadequately performed Valsalva maneuver. The resulting 20 patients showed a clear increase in oxygenated and deoxygenated hemoglobin concentration after the onset of the Valsalva maneuver. ROC analysis yielded AUC values (0.393 - 0.779) that did not differ from random probabilities. The highest AUC values were obtained for FWHM (AUC: 0.779, detection rates [60 - 70 %], identification of additional lesions [55 - 70 %]). PA analysis had the highest detection rate (70 - 90 %) but also the highest identification of false-positive additional lesions (80 - 90 %). The concordance rates of the two readers for malignant lesions were satisfactory (0.524 - 1.0). Our study revealed susceptibility to artifacts and a large number of false-positive additional lesions, suggesting that the evaluation of hemodynamic changes after Valsalva maneuver by optical imaging is not a promising method.


Risk of hepatitis B virus reactivation in rheumatoid arthritis patients undergoing biologic treatment: Extending perspective from old to newer drugs.
Hepatitis B virus (HBV) reactivation in rheumatoid arthritis (RA) patients undergoing biological therapy is not infrequent. This condition can occur in patients with chronic hepatitis B as well as in patients with resolved HBV infection. Current recommendations are mainly focused on prevention and management strategies of viral reactivation under tumor necrosis factor-α inhibitors or chimeric monoclonal antibody rituximab. In recent years, growing data concerning HBV reactivation in RA patients treated with newer biological drugs like tocilizumab and abatacept have cumulated. In this review, epidemiology, pathogenesis and natural history of HBV infection have been revised first, mainly focusing on the role that specific therapeutic targets of current biotechnological drugs play in HBV pathobiology; finally we have summarized current evidences from scientific literature, including either observational studies and case reports as well, concerning HBV reactivation under different classes of biological drugs in RA patients. Taking all these evidences into account, some practical guidelines for screening, vaccination, prophylaxis and treatment of HBV reactivation have been proposed.


Effects of concurrent vaginal miconazole treatment on the absorption and exposure of Nestorone® (segesterone acetate) and ethinyl estradiol delivered from a contraceptive vaginal ring: a randomized, crossover drug-drug interaction study.
To evaluate the effects of concurrent administration of three vaginal miconazole nitrate formulations on the absorption and exposure of Nestorone® (segesterone acetate) and ethinyl estradiol from a novel contraceptive vaginal ring (NES/EE CVR). This was an open-label, randomized, crossover, drug-drug interaction study conducted over three menstrual cycles in healthy women with regular menses. We compared systemic exposure to NES and EE by determining area under the curve (AUC8-21d) with CVR only and CVR with each miconazole treatment. Three different miconazole formulations (single-dose suppository, multiple-dose suppository or multiple-dose cream) were administered in a single dose on day 8 or multiple doses on days 8-10 after CVR insertion. We evaluated safety and tolerability of the CVR in the presence of antimycotic comedication. Forty-five participants were randomized, and 29 completed participation. Systemic exposure to NES and EE released from the CVR increased with single or multiple doses of miconazole suppositories but not with multiple-dose cream. The maximum EE geometric mean ratio (GMR) for AUC8-21d was 1.67 (1.51-1.86) for single-dose and 1.42 (1.21-1.66) for multiple-dose suppositories. By contrast, systemic exposure to NES and EE was comparable with and without miconazole cream (all GMRs and confidence intervals within 0.80 to 1.25). Adverse events (AEs) were similar with CVR only and with all miconazole treatment groups. There were no serious treatment-related AEs. Miconazole vaginal suppositories were associated with increased systemic levels of NES and EE, while systemic exposure with miconazole vaginal cream was comparable to no miconazole exposure. Coadministration of miconazole suppositories with the investigational NES/EE CVR led to higher systemic exposure of both hormones, while coadministration with miconazole cream did not affect hormone levels. Women utilizing the NES/EE CVR may be advised to use an oral formulation or miconazole cream rather than suppository to treat vaginal candidiasis.


The metastatic microenvironment: Brain-derived soluble factors alter the malignant phenotype of cutaneous and brain-metastasizing melanoma cells.
The working hypothesis of this study is that the interactions between the brain microenvironment and melanoma cells determine metastasis formation at this organ site. The aim of the study was to evaluate the contribution of such interactions to the formation of brain metastasis in nude mice xenografted with human melanoma cells. An insight into these interactions is an essential prerequisite for the development of effective targeted therapy for melanoma brain metastasis. We assessed the effects of soluble factors present in supernatants of short-term cultures of normal mouse brain (referred here after as brain-derived soluble factors) on several characteristics linked to melanoma brain metastasis. It was found that brain-derived soluble factors affect differentially cutaneous and brain-metastasizing melanoma cells variants in vitro. Such factors enhanced the viability of cutaneous melanoma cells but caused an S phase arrest followed by apoptosis of brain-metastasizing cells. Brain-derived soluble factors enhanced migration of melanoma cells metastasizing to the brain, but did not affect the migration of the cutaneous variants. Such factors upregulated the expression of the chemokine receptor CCR4 in both cutaneous and brain-metastasizing melanoma cells. It is not unlikely that CCR4 ligands expressed in the brain interact with the CCR4-expressing melanoma cells thereby directing them to the brain. Brain-derived soluble factors enhanced the transmigration, across human brain endothelial cells of cutaneous but not of brain-metastasizing melanoma variants. This activity could promote the capacity of the cutaneous cells to metastasize to the brain.


New developments in 'any willing provider' laws.
As the debate over healthcare reform continues, the controversy concerning "any willing provider" laws also continues. "Any willing provider" laws require a healthcare insurer (or managed care organization) to establish criteria that a provider must meet in order to participate as a network provider. Such laws prohibit an insurer from denying any provider willing to meet those criteria from participating as a network provider. Proponents of any willing provider laws cite the recent decision by the U.S. Supreme Court not to review a lower court ruling finding in favor of a hospital as validation of such laws.


Calmodulin binding to the dehydrogenase domain of NADPH oxidase 5 alters its oligomeric state.
Superoxide generated by NADPH Oxidase 5 (Nox5) is regulated by Ca2+ through the interaction of its self-contained Ca2+ binding domain and dehydrogenase domain (DH). Recently, calmodulin (CaM) has been reported to enhance the Ca2+ sensitivity of Nox5 by binding to the CaM-binding domain sequence (CMBD), in which the interaction between CaM and Nox5 is largely unclear. Here, we used the CMBD peptide and truncated DH constructs, and separately studied their interaction with CaM by fluorescence, calorimetry, and dynamic light scattering. Our results revealed that each half-domain of CaM binds one CMBD peptide with a binding constant near 106 M-1 and a binding enthalpy change of -3.81 kcal/mol, consistent with an extended 1:2 CaM:CMBD structure. However, the recombinant truncated DH proteins exist as oligomers, possibly trimer and tetramer. The oligomeric states are concentration and salt dependent. CaM binding appears to stabilize the DH dimer complexed with CaM. The thermodynamics of CaM binding to the DH is comparable to the peptide-based study except that the near unity binding stoichiometry and a large conformational change were observed. Our result suggests that the oligomeric states of Nox5, mediated by its DH domain and CaM, may be important for its superoxide-generating activity.


Effects of age and body mass index on the results of transtrochanteric rotational osteotomy for femoral head osteonecrosis.
Advanced-stage osteonecrosis and a large area of necrotic bone are known risk factors for failure of transtrochanteric rotational osteotomy of the hip in patients with osteonecrosis. The purpose of this study was to determine whether there are other risk factors for failure of this osteotomy. One hundred and five patients (113 hips) underwent an anterior transtrochanteric rotational osteotomy for the treatment of femoral head osteonecrosis and were followed for a mean of 51.3 months postoperatively. Radiographic failure was defined as secondary collapse or osteoarthritic change. Multivariate analysis was performed to assess factors associated with secondary collapse and osteophyte formation. The Kaplan-Meier product-limit method was used to estimate survival. Secondary collapse occurred in twenty-seven hips (24%), and fourteen hips (12%) were converted to a total hip arthroplasty. At the time of the most recent follow-up, the hip scores according to the system of Merle d'Aubigné et al. ranged from 6 to 18 points (mean, 15.8 points). Multivariate analysis showed that the stage of the necrosis (III or greater) (hazard ratio = 3.28; 95% confidence interval = 1.49 to 7.24), age of the patient (forty years or older) (hazard ratio = 1.08; 95% confidence interval = 1.02 to 1.14), body mass index (> or = 24 kg/m(2)) (hazard ratio = 1.19; 95% confidence interval = 1.03 to 1.38), and extent of the necrosis (a combined necrotic angle of > or = 230 degrees ) (hazard ratio = 1.08; 95% confidence interval = 1.04 to 1.11) were associated with secondary collapse. Seven of the eighty-six hips without collapse showed progression to osteoarthritis. The survival rate at 110 months was 63.4% (95% confidence interval = 51.1% to 75.7%) with total hip arthroplasty or radiographic failure as the end point and 56.0% (95% confidence interval = 44.6% to 67.4%) with total hip arthroplasty, radiographic failure, or loss to follow-up as the end point. Our study showed that age, body mass index, and the stage and extent of the osteonecrosis were determining factors for secondary collapse, unsatisfactory clinical results, and conversion to total hip arthroplasty. These factors should be considered when selecting patients for a transtrochanteric rotational osteotomy.


Solver Device for Powdery Drugs.
Pharmacotherapy is a major treatment method in healthcare centers, and the injection of powdered drugs is among common pharmacotherapy techniques. Medication errors and nosocomial infections are among major health issues in the world. On the other hand, powdered drugs are widely used in hospitals; however, drug mixture is a very time-consuming process. The objective of this invention was to accelerate drug vial mixture, reduce medication errors and nosocomial infections, and save time. There are different drug mixing devices, each with specific abilities. The present invention not only possesses the abilities of other devices but also can mix drugs [with a diluent] with higher quality and accuracy. The drug vial mixing device can mix 20 vials at adjustable time and speed with high quality and accuracy. This device is equipped with an infrared system to ensure a complete mixture of powder, and a Bluetooth short range radio system to remotely control all monitoring options of the device. This is a small and simple-to-use device. The drug vial mixing device can effectively reduce medication errors and nosocomial infections, and save time.


Negative co-operativity in glutamate dehydrogenase. Involvement of the 2-position in glutamate in the induction of conformational changes.
The 2-position substituent on substrates or substrate analogues for glutamate dehydrogenase is shown to be intimately involved in the induction of conformational changes between subunits in the hexamer by coenzyme. These conformational changes are associated with the negative co-operativity exhibited by this enzyme. 2-Oxoglutarate and L-2-hydroxyglutarate induce indications of co-operativity similar to those induced by the substrate of oxidative deamination, glutamate, in kinetic studies. Glutarate (2-position CH2) does not. A comparison of the effects of L-2-hydroxyglutarate and D-2-hydroxyglutarate or D-glutamate indicates that the 2-position substituent must be in the L-configuration for these conformational changes to be triggered. In addition, glutarate and L-glutamate in ternary enzyme-NAD(P)H-substrate complexes induce very different coenzyme fluorescence properties, showing that glutamate induces a different conformation of the enzyme-coenzyme complex from that induced by glutarate. Although glutamate and glutarate both tighten the binding of reduced coenzyme to the active site, the effect is much greater with glutamate, and the binding is described by two dissociation constants when glutamate is present. The data suggest that the two carboxy groups on the substrate are required to allow synergistic binding of coenzyme and substrate to the active site, but that interactions between the 2-position on the substrate and the enzyme trigger the conformational changes that result in subunit-subunit interactions and in the catalytic co-operativity exhibited by this enzyme.


Pharmacologic disposition of inosine dialdehyde (NSC-118994) in mice, rats, dogs, and monkeys.
The pharmacologic disposition of inosine dialdehyde (IdA) was studied in mice, rats, dogs, and monkeys. 14C-IdA was utilized to follow plasma and tissue levels and patterns of excretion. Plasma radioactivity curves were multiphasic with an initial rapid component (half-life = 12-18 mins) followed by a much slower decline (half-life = approximately 3 days). As time progressed after the injection, an increasing fraction of the total plasma radioactivity was found to be associated with plasma proteins and was trichloracetic acid (TCA) insoluble. IdA interacts extensively with proteins and small molecules both in vivo and in vitro, forming complexes that do not dissociate under physiologic conditions. The pharmacologic disposition of IdA is dependent upon the relative extent of IdA interaction with protein macromolecules versus small transportable molecules. The major route of excretion was urinary with less than 50% of the dose excreted during the first 24 hours after iv administration in all species studied. IdA was recovered in the urine as complexes with urine constituents. There was no preferential uptake of IdA by any organ. All radioactivity remaining 24 hours after IdA administration was associated with plasma and cellular proteins and was nondiayzable and TCA insoluble. IdA was not a substrate for aldehyde oxidase and did not inhibit the activity of this enzyme.


Nuclear DNA content and chromatin pattern of rat rhabdomyosarcoma cell sublines with different metastatic potentials.
There is a constant need of features able to characterize potentially metastatic cells among the heterogeneous cell subpopulations which constitute a tumor. Image cytometry of metastatic tumor cells give rise to variable results, partly because of a heterogeneous origin of cells, or potential drug effects. The aim of this work was to characterize nuclear changes observed in metastatic cell clones issued in vitro from the same parental cell population The nuclear phenotypes of 6 cell sublines isolated from a rat rhabdomyosarcoma cell line and differing in their metastatic ability were evaluated by image cytometry on Feulgen-stained preparations. Densitometric [5], geometric [3] and textural [9] features were computed from each nuclear image. For each cell subline, a metastatic score, ranging from 0 to 10, was calculated on the basis of in vivo invasivity data, by measuring the number of pulmonary metastases observed after s.c. graft of tumor cells in rats. Data obtained were compared to karyotype, growth characteristics, and oncogene expressions of cell lines. The nuclear DNA content, the chromosome numbers, the cell sublines doubling times, and the distribution of cells within the cell cycle appear unrelated with this score. On the contrary, increase in metastatic ability is accompanied by changes in chromatin pattern as assessed by textural features. Progressive increase in chromatin condensation can be observed in cell sublines with increasing metastatic score. These results were confirmed by an unsupervised multivariate partitioning of rhabdomyosarcoma cells which identified two separate subsets whose distributions within the analyzed cell lines correlate with their metastatic ability. These data suggest that, in rat rhabdomyosarcoma cell sublines, metastatic ability could be associated with nuclear morphological changes at the level of chromatin texture.


Clinical observations on smallpox: a study of 1233 patients admitted to the Infectious Diseases Hospital, Calcutta, during 1973.
The paper presents clinical observations on 1 233 persons with smallpox who were admitted to the Infectious Diseases Hospital, Calcutta, in 1973. The disease was of the modified type in 53 patients (4.3%), the ordinary type in 717 (58.2%), the flat type in 249 (20.2%), and the haemorrhagic type in 214 (17.3%). The fatality of these types of smallpox was found to be 5.7%, 26.8%, 88.4%, and 98.1%, respectively, and the overall case fatality was 50.7%. The haemorrhagic type was found mainly among older patients and affected males more often than females. The vaccination status of 1 218 patients was known. Of these, 901 (73.9%) were unvaccinated and had a fatality rate of 53.4%, whereas the 317 (26.1%) vaccinated patients had a fatality rate of 36.5%. Among the 201 haemorrhagic cases, 145 patients were unvaccinated (16.09% of the total number unvaccinated) and 56 (17.67%) had been vaccinated. Of 34 patients vaccinated during the incubation period, 19 (41.1%) died, whereas of 18 patients who had been vaccinated after the onset of fever, but before the appearance of rash, 9 (50%) died.


Potency Ranking of Per- and Polyfluoroalkyl Substances Using High-Throughput Transcriptomic Analysis of Human Liver Spheroids.
Per- and polyfluoroalkyl substances (PFAS) are some of the most prominent organic contaminants in human blood. Although the toxicological implications of human exposure to perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) are well established, data on lesser-understood PFAS are limited. New approach methodologies (NAMs) that apply bioinformatic tools to high-throughput data are being increasingly considered to inform risk assessment for data-poor chemicals. The aim of this study was to compare the potencies (ie, benchmark concentrations: BMCs) of PFAS in primary human liver microtissues (3D spheroids) using high-throughput transcriptional profiling. Gene expression changes were measured using TempO-seq, a templated, multiplexed RNA-sequencing platform. Spheroids were exposed for 1 or 10 days to increasing concentrations of 23 PFAS in 3 subgroups: carboxylates (PFCAs), sulfonates (PFSAs), and fluorotelomers and sulfonamides. PFCAs and PFSAs exhibited trends toward increased transcriptional potency with carbon chain-length. Specifically, longer-chain compounds (7-10 carbons) were more likely to induce changes in gene expression and have lower transcriptional BMCs. The combined high-throughput transcriptomic and bioinformatic analyses support the capability of NAMs to efficiently assess the effects of PFAS in liver microtissues. The data enable potency ranking of PFAS for human liver cell spheroid cytotoxicity and transcriptional changes, and assessment of in vitro transcriptomic points of departure. These data improve our understanding of the possible health effects of PFAS and will be used to inform read-across for human health risk assessment.


Experimental metabolic surgery: justification and technical aspects.
Metabolic surgery is a surgical strategy which has shown great potential in the treatment of diseases which may be associated with morbid obesity. It must be developed on the basis of both animal and clinical research. The objective of this study is to set out the various options in experimentation animals and the technical characteristics in operations, and the specific animal care undertaken by our group. We identified and reviewed the key points to be considered in animal handling during interventions such as sleeve gastrectomy, Roux-en-Y gastric bypass, ileal transposition and duodenal exclusion. The technical variations found at experimental level are due to the pouch capacity for the Roux-en-Y gastric bypass. Intestinal anastomosis is the variable with the greatest differences found between the various working groups. Ileal transposition is a technique that is undergoing constant review, and the results differ substantially depending on the animal model chosen, and are also metabolically effective in animals with a normal weight. Duodenal exclusion by means of a physical barrier has not been studied sufficiently but could be a pre-operative support for weight loss. There are experimental technical discrepancies and further studies are necessary to ascertain their efficiency. Metabolic surgery currently complements bariatric surgery and justifies the appearance of new experimental studies. The animal models chosen are very important as only very specific study models will be used in cases in which the technique is sufficiently validated by the research team, as the results to be assessed depend on this.


Impact of asymptomatic herpes simplex virus type 2 infection on mucosal homing and immune cell subsets in the blood and female genital tract.
HSV-2 infection is common and generally asymptomatic, but it is associated with increased HIV susceptibility and disease progression. This may relate to herpes-mediated changes in genital and systemic immunology. Cervical cytobrushes and blood were collected from HIV-uninfected African/Caribbean women in Toronto, and immune cell subsets were enumerated blindly by flow cytometry. Immune differences between groups were assessed by univariate analysis and confirmed using a multivariate model. Study participants consisted of 46 women, of whom 54% were infected with HSV-2. T cell activation and expression of the mucosal homing integrin α4β7 (19.60 versus 8.76%; p < 0.001) were increased in the blood of HSV-2-infected women. Furthermore, expression of α4β7 on blood T cells correlated with increased numbers of activated (coexpressing CD38/HLA-DR; p = 0.004) and CCR5(+) (p = 0.005) cervical CD4(+) T cells. HSV-2-infected women exhibited an increase in the number of cervical CD4(+) T cells (715 versus 262 cells/cytobrush; p = 0.016), as well as an increase in the number and proportion of cervical CD4(+) T cells that expressed CCR5(+) (406 versus 131 cells, p = 0.001; and 50.70 versus 34.90%, p = 0.004) and were activated (112 versus 13 cells, p < 0.001; and 9.84 versus 4.86%, p = 0.009). Mannose receptor expression also was increased on cervical dendritic cell subsets. In conclusion, asymptomatic HSV-2 infection was associated with significant systemic and genital immune changes, including increased immune activation and systemic α4β7 expression; correlation of the latter with highly HIV-susceptible CD4(+) T cell subsets in the cervix may provide a mechanism for the increased HIV susceptibility observed in asymptomatic HSV-2-infected women.


Serum S100B Protein is Specifically Related to White Matter Changes in Schizophrenia.
Schizophrenia can be conceptualized as a form of dysconnectivity between brain regions.To investigate the neurobiological foundation of dysconnectivity, one approach is to analyze white matter structures, such as the pathology of fiber tracks. S100B is considered a marker protein for glial cells, in particular oligodendrocytes and astroglia, that passes the blood brain barrier and is detectable in peripheral blood. Earlier Studies have consistently reported increased S100B levels in schizophrenia. In this study, we aim to investigate associations between S100B and structural white matter abnormalities. We analyzed data of 17 unmedicated schizophrenic patients (first and recurrent episode) and 22 controls. We used voxel based morphometry (VBM) to detect group differences of white matter structures as obtained from T1-weighted MR-images and considered S100B serum levels as a regressor in an age-corrected interaction analysis. S100B was increased in both patient subgroups. Using VBM, we found clusters indicating significant differences of the association between S100B concentration and white matter. Involved anatomical structures are the posterior cingulate bundle and temporal white matter structures assigned to the superior longitudinal fasciculus. S100B-associated alterations of white matter are shown to be existent already at time of first manifestation of psychosis and are distinct from findings in recurrent episode patients. This suggests involvement of S100B in an ongoing and dynamic process associated with structural brain changes in schizophrenia. However, it remains elusive whether increased S100B serum concentrations in psychotic patients represent a protective response to a continuous pathogenic process or if elevated S100B levels are actively involved in promoting structural brain damage.


Management of major trauma: changes required for improvement.
To describe the views of key healthcare professionals on the changes they considered to be important in the reduction of major trauma mortality between 1988 and 1995 in Leeds. Qualitative unstructured interviews with a purposive sample of 10 healthcare professionals deemed to be key personnel by an experienced consultant who had provided acute trauma care throughout the relevant period. Each interview was tape recorded and transcribed; each transcript was analysed for important themes by two independent researchers who then discussed their results to resolve any differences in interpretation. Three categories of change became evident: "policy", "infrastructure", and "philosophy of care". Each of these categories seemed to be equally important. Policy changes identified as important were the Royal College of Surgeons of England's report into trauma care (1988), the setting of standards for paramedic training, and the national audit of major trauma outcomes. Important infrastructure changes identified were training in advanced trauma life support, decreased ambulance response times, reorganisation towards "consultant led" hospital services, and an emphasis on quality monitoring. Changes in philosophy of care were increases in levels of teamwork, commitment, communication, and confidence. Together these facilitated an overall restructuring and refocusing of care. No individual change is seen as dominant for improved care, but rather a strategic mixture of facilitating national and regional policy guidance, organisational restructuring, and congruent professional attitudes were integral components leading to the observed changes. Improving outcomes in other areas is likely to involve an integrated series of changes which must be managed as a total system.


Morphometric analysis of thyroid gland in adult north Indian population by nuclear imaging.
A detailed retrospective morphometric analysis of thyroid gland in adult north Indian subjects using scintillation imaging has been done. Many parameters were found to be significantly different as compared to non-Indian population. Utility of this analysis has been discussed.


Using medical knowledge sources on handheld computers--a qualitative study among junior doctors.
The emergence of mobile computing could have an impact on how junior doctors learn. To exploit this opportunity it is essential to understand their information seeking process. To explore junior doctors' experiences of using medical knowledge sources on handheld computers. Interviews with five Swedish junior doctors. A qualitative manifest content analysis of a focus group interview followed by a qualitative latent content analysis of two individual interviews. A focus group interview showed that users were satisfied with access to handheld medical knowledge sources, but there was concern about contents, reliability and device dependency. Four categories emerged from individual interviews: (1) A feeling of uncertainty about using handheld technology in medical care; (2) A sense of security that handhelds can provide; (3) A need for contents to be personalized; (4) A degree of adaptability to make the handheld a versatile information tool. A theme was established to link the four categories together, as expressed in the Conclusion section. Junior doctors' experiences of using medical knowledge sources on handheld computers shed light on the need to decrease uncertainty about clinical decisions during medical internship, and to find ways to influence the level of self-confidence in the junior doctor's process of decision-making.


Dose response and atypical antipsychotics in schizophrenia.
Based on information from clinical trials, both the efficacy and adverse effects of conventional antipsychotics in the treatment of schizophrenia are dose related. The overlapping nature of these dose-response profiles limits the use of these agents. Atypical antipsychotics provide greater relief across the comorbid symptom domains of schizophrenia, but dose-response studies and clinical experience have revealed that some of these drugs also have dose limitations. This article reviews the dose-response relationships of the atypical antipsychotics as presented predominantly in pivotal, randomised studies (double-blind and otherwise). Limited data indicate that clozapine shows dose-related efficacy up to 600 mg/day in patients with treatment-resistant schizophrenia. However, higher dosages of clozapine may be associated with the risk of seizures. Risperidone demonstrates dose-related adverse events that compromise efficacy. The dose-response relationships for ziprasidone, quetiapine and aripiprazole are less well established. The efficacy of olanzapine appears to be dose related within the recommended dosage range of 10-20 mg/day, but clinical trials that have explored higher dosages suggest improved efficacy. Furthermore, the higher doses are not associated with a significantly increased incidence of adverse events. Further studies are clearly needed to fully characterise the dose-response relationships of atypical antipsychotics.


Weaning children from mechanical ventilation with a computer-driven protocol: a pilot trial.
Duration of weaning from mechanical ventilation is decreased with the use of written protocols in adults. In children, the use of written protocols has not had such an impact. We conducted a single-center trial to assess the feasibility of conducting a multicenter randomized clinical trial comparing the duration of weaning from mechanical ventilation in those managed by a computer-driven explicit protocol versus usual care. Mechanically ventilated children aged between 2 and 17 years on pressure support and not receiving inotropes were included. After randomization, children were weaned either by usual care (n = 15) that was characterized by no protocolized decisions by attending physicians, or by a computer-driven protocol (Smartcare/PS™, Drager Medical) (n = 15). Weaning duration until first extubation was the primary outcome. For comparison, a Mann-Whitney U test was employed (p < 0.05). Patients characteristics at inclusion were similar. The median duration of weaning was 21 h (range 3-142 h) in the SmartCare/PS™ group and 90 h (range 4-552 h) in the usual care group, p = 0.007. The rate of reintubation within 48 h after extubation and the rate of noninvasive ventilation after extubation in the SmartCare/PS™ and usual care groups were 2/15 versus 1/15 and 2/15 versus 2/15, respectively. A pediatric randomized trial on mechanical ventilation with a computerized protocol in North America is feasible. A computer-driven protocol that also manages children younger than 2 years old would help to decrease the number of PICU admissions screened in a multicentre trial on this topic.


Eating Disorders and Disordered Eating in Competitive Cycling: A Scoping Review.
This article reports on the findings from a scoping review on eating disorders and disordered eating in competitive cycling. The review was informed by a scoping review methodological framework as well as the Preferred Reporting Items for Systematic Reviews and Meta-analysis extension for scoping reviews (PRISMA-ScR) reporting guidelines. PubMed, SPORTDiscus and Web of Science were used to identify relevant literature for review. Fourteen studies met the eligibility criteria and were included in the full review. A narrative synthesis was used to summarise the main findings and themes across the included literature. Findings from the review are presented under the following themes: cycling as an 'at-risk' discipline; power to weight ratio; energy requirements and risk of low energy availability; the social environment of cycling; nutrition support provision; relationship between eating disorders/disordered eating and exercise addiction; and recommendations made in identified literature. Overall, the literature suggests competitive cycling is a sport with a high prevalence of disordered eating and/or eating disorders and a sport with unique risk factors that contribute to this. Crucially, more research is needed in this area. The article concludes with the gaps in the literature highlighted, implications for future research, and applications to policy and practice suggested.


Tolerance and compensatory response of rice to sugarcane borer (Lepidoptera: Crambidae) injury.
A 3-yr field experiment was conducted to evaluate the tolerance and compensatory response of rice (Oryza sativa L.) to injury caused by sugarcane borer, Diatraea saccharalis (F.), as affected by cultivar (Cocodrie, Francis, and Jefferson), stage of crop growth during which the injury occurred (third tiller stage, panicle differentiation stage, and heading stage), and sugarcane borer density. The proportion of rice tillers with sugarcane borer injury (leaf and leaf sheath injury and/or stem injury) was lower when injury occurred at the third tiller stage (0.05) than at panicle differentiation (0.19) and heading (0.18). When injury occurred at the two latter stages, both the proportion of tillers with injury and the proportion of tillers with stem injury were negatively correlated with rainfall. Rainfall resulted in dislodgement and mortality of sugarcane borer eggs and larvae before the larvae entered the stems. Rice plant density in this study (111.1 plants/m2) was higher than recorded for previous research on rice compensation using potted rice or conducted in low-density hill production systems (26.7-51.3 plants/m2). Two mechanisms of within-plant tolerance/compensation were observed. Stem injured plants produced approximately 0.69 more tillers than uninjured plants, whereas tillers with leaf and leaf sheath injury produced larger panicles, up to 39.5 and 21.0% heavier than uninjured tillers, when injury occurred at third tiller stage and at panicle differentiation, respectively. Rice yield was not reduced with up to 23% injured tiller and up to 10% injured stems at the third tiller stage, 42% injured tillers and 17% injured stems at panicle differentiation, and 28% injured tillers and 14% injured stems at heading. Significant between-plant compensation was not detected, suggesting competition between adjacent plants is not significantly reduced by injury. Our results suggest that rice can tolerate and/or compensate for a level of stem borer injury previously considered to be economically damaging.


Cancer screening of long-term cancer survivors.
Although 64% of cancer survivors are expected to live at least 5 years beyond diagnosis, the receipt of cancer screening by this population is unclear. The objective of this study was to assess the relation between a cancer diagnosis and future cancer screening, exploring provider-, patient-, and cancer-specific factors that explain observed relationships. The Wisconsin Longitudinal Study (WLS) and Wisconsin Tumor Registry were used to identify 2 participant groups: 415 patients diagnosed with nonmetastatic cancer between 1992 to 1993 (before cancer) and 2003 to 2004 (after cancer) and 4680 controls (no cancer). Adjusted average predicted probabilities of cancer screening were estimated with models that first did not include and then included, provider (provider relationship length), participant (depressive symptoms per the Center for Epidemiologic Studies Depression Scale), and cancer-specific (time since diagnosis) factors. Participants with a history of cancer associated with a given screening test were then excluded to assess whether relationships are explained by screening for recurrence versus second cancers. Female cancer survivors were more likely than no-cancer controls to undergo pelvic/Papanicolaou screening (survivors: 70%, 95% confidence interval [CI]: 63% to 76%; controls: 61%, 95% CI: 59% to 63%) and mammography screening (survivors: 86%, 95% CI: 78% to 90%; controls: 76%, 95% CI: 74% to 77%), though male cancer survivors were not more likely to receive prostate exams (survivors: 76%, 95% CI: 70% to 82%; controls: 69%, 95% CI: 67% to 71%). After excluding people with a history of the cancer being screened for, there were few significant differences in cancer screening between short- or long-term survivors (≥5 years) and no-cancer controls. Relationships were not sensitive to adjustment for provider or participant factors. The significant positive differences in cancer screening between people with and without cancer can be explained by screening for recurrence. Long-term cancer survivors are not more likely to receive follow-up screening for second cancers. This information should be used by providers to ensure patients receive recommended follow-up preventive care.


Novel genes involved in the regulation of pathogenicity factor production within the rpf gene cluster of Xanthomonas campestris.
The synthesis of extracellular enzymes and extracellular polysaccharide (EPS) in Xanthomonas campestris pathovar campestris (Xcc) is subject to co-ordinate regulation by a cluster of genes called rpf (for regulation of pathogenicity factors). These genes are located within a 21.9 kb region of the chromosome isolated as the cosmid clone pIJ3020. The genes in the left-hand section of this region of the chromosome have previously been characterized. This paper reports on the genes in the right-hand section and on the phenotypes of mutants with transposon insertions in these genes. Sequence analysis identified eight genes or ORFs with the gene order rpfD-orf1-orf2-orf3-orf4-recJ-rpf E-greA. RecJ and GreA have established functions in recombination and transcriptional elongation, respectively. rpfD encoded a protein with some amino acid sequence relatedness to a hypothetical protein from Caulobacter crescentus and an autolysin response regulator in Bacillus subtilis. The predicted protein products of orf1, 2 and 3 were related to each other and had substantial amino acid sequence relatedness to hypothetical proteins from C. crescentus. Transposon insertions in orf1, 2 and 3 had no effect on the synthesis of extracellular enzymes or EPS. The predicted proteins RpfE and Orf4 showed the highest amino acid sequence relatedness to hypothetical proteins from Bordetella pertussis and Klebsiella pneumoniae, respectively. Transposon insertions in rpfE led to reduced levels of some extracellular enzymes (endoglucanase and protease) and increased levels of others (polygalacturonate lyase). Transposon insertions in orf4 had no effect on polygalacturonate lyase but led to reduced levels of protease and endoglucanase. Levels of EPS were reduced in both rpfE and orf4 mutants. These alterations in the levels of extracellular enzymes, which were relatively modest (between two- and threefold), did not affect the pathogenicity of Xcc on turnip. It is proposed that the gene designation should be rpfI for orf4.


FISH of a maize sh2-selected sorghum BAC to chromosomes of Sorghum bicolor.
Fluorescence in situ hybridization (FISH) of a 205 kb Sorghum bicolor bacterial artificial chromosome (BAC) containing a sequence complementary to maize sh2 cDNA produced a large pair of FISH signals at one end of a midsize metacentric chromosome of S. bicolor. Three pairs of signals were observed in metaphase spreads of chromosomes of a sorghum plant containing an extra copy of one arm of the sorghum chromosome arbitrarily designated with the letter D. Therefore, the sequence cloned in this BAC must reside in the arm of chromosome D represented by this monotelosome. This demonstrates a novel procedure for physically mapping cloned genes or other single-copy sequences by FISH, sh2 in this case, by using BACs containing their complementary sequences. The results reported herein suggest homology, at least in part, between one arm of chromosome D in sorghum and the long arm of chromosome 3 in maize.


Mutations in SREBF1, Encoding Sterol Regulatory Element Binding Transcription Factor 1, Cause Autosomal-Dominant IFAP Syndrome.
IFAP syndrome is a rare genetic disorder characterized by ichthyosis follicularis, atrichia, and photophobia. Previous research found that mutations in MBTPS2, encoding site-2-protease (S2P), underlie X-linked IFAP syndrome. The present report describes the identification via whole-exome sequencing of three heterozygous mutations in SREBF1 in 11 unrelated, ethnically diverse individuals with autosomal-dominant IFAP syndrome. SREBF1 encodes sterol regulatory element-binding protein 1 (SREBP1), which promotes the transcription of lipogenes involved in the biosynthesis of fatty acids and cholesterols. This process requires cleavage of SREBP1 by site-1-protease (S1P) and S2P and subsequent translocation into the nucleus where it binds to sterol regulatory elements (SRE). The three detected SREBF1 mutations caused substitution or deletion of residues 527, 528, and 530, which are crucial for S1P cleavage. In vitro investigation of SREBP1 variants demonstrated impaired S1P cleavage, which prohibited nuclear translocation of the transcriptionally active form of SREBP1. As a result, SREBP1 variants exhibited significantly lower transcriptional activity compared to the wild-type, as demonstrated via luciferase reporter assay. RNA sequencing of the scalp skin from IFAP-affected individuals revealed a dramatic reduction in transcript levels of low-density lipoprotein receptor (LDLR) and of keratin genes known to be expressed in the outer root sheath of hair follicles. An increased rate of in situ keratinocyte apoptosis, which might contribute to skin hyperkeratosis and hypotrichosis, was also detected in scalp samples from affected individuals. Together with previous research, the present findings suggest that SREBP signaling plays an essential role in epidermal differentiation, skin barrier formation, hair growth, and eye function.


Old yeasts, young beer-The industrial relevance of yeast chronological life span.
Much like other living organisms, yeast cells have a limited life span, in terms of both the maximal length of time a cell can stay alive (chronological life span) and the maximal number of cell divisions it can undergo (replicative life span). Over the past years, intensive research revealed that the life span of yeast depends on both the genetic background of the cells and environmental factors. Specifically, the presence of stress factors, reactive oxygen species, and the availability of nutrients profoundly impact life span, and signaling cascades involved in the response to these factors, including the target of rapamycin (TOR) and cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) pathways, play a central role. Interestingly, yeast life span also has direct implications for its use in industrial processes. In beer brewing, for example, the inoculation of finished beer with live yeast cells, a process called "bottle conditioning" helps improve the product's shelf life by clearing undesirable carbonyl compounds such as furfural and 2-methylpropanal that cause staling. However, this effect depends on the reductive metabolism of living cells and is thus inherently limited by the cells' chronological life span. Here, we review the mechanisms underlying chronological life span in yeast. We also discuss how this insight connects to industrial observations and ultimately opens new routes towards superior industrial yeasts that can help improve a product's shelf life and thus contribute to a more sustainable industry.


A biomechanical analysis of intravertebral pressures during vertebroplasty of cadaveric spines with and without simulated metastases.
A biomechanical cadaveric study of thoracic and lumbar vertebrae with simulated metastases quantifying intravertebral pressures during transpedicular vertebroplasty. To compare intravertebral pressures during percutaneous vertebroplasty in vertebrae with and without simulated lytic metastases. Percutaneous vertebroplasty is designed to provide stability to vertebrae weakened by osteoporosis or metastatic disease. The complication rate is higher when the procedure is used for the treatment of lytic vertebral lesions. The major complications reported are radiculopathy, spinal cord compression, and embolic phenomena. Ten fresh-frozen cadaveric vertebrae were tested intact (7 lumbar, 3 thoracic) and 7 were tested with simulated lytic defects (4 lumbar, 3 thoracic). Defects were created by replacing a core of cancellous bone with soft tumor tissue in the center of the vertebral body. Simplex P (Howmedica Osteonics, Mahwah, NJ) cement was injected into each vertebra through a unipedicular approach at a constant rate of 3 mL per minute. Cement volume, injection force, and intravertebral pressures at the posterior vertebral body wall were recorded. Following the procedure, the vertebrae were sectioned to visualize cement and tumor disbursement. There was no significant difference between the two groups for age, size, trabecular density, and cement volume. Vertebrae with simulated metastases generated an average maximum pressure of 39.66 kPa during cement injection versus 6.83 kPa in intact vertebrae (P < 0.05). Higher pressures were also generated in smaller vertebrae based on a power relationship (r2 = 0.71 intact, r2 = 0.43 tumor). Percutaneous vertebroplasty produces higher intravertebral pressures in vertebrae containing a simulated lytic metastasis than in intact vertebrae. Pressures generated in the tumor specimens are sufficiently elevated to cause embolic phenomena.


Substrate adhesion of rat hepatocytes: a comparison of laminin and fibronectin as attachment proteins.
In previous studies rat hepatocytes have been shown to adhere to substrates composed of collagen or fibronectin. In the present communication, the basement membrane protein laminin is reported to mediated the attachment and spreading of hepatocytes. The cell attachment-mediating activity of laminin was compared with that of fibronectin. The activity of fibronectin was heat sensitive, whereas laminin retained its activity after boiling. On the other hand, reduction and alkylation or periodate oxidation of the proteins affected only the cell attachment activity of laminin. Preincubation of cells with soluble fibronectin inhibited initial cell attachment to fibronectin but not to laminin substrates, and, reversely, soluble laminin selectively inhibited cell attachment to laminin. These results suggest that attachment of cells to substrates of the two proteins involves different cellular receptors recognizing distinct and nonidentical structures in the proteins.


Epithelioid trophoblastic tumor of uterus presenting as an ovarian mass: a diagnostic and therapeutic dilemma.
Epithelioid trophoblastic tumor (ETT) is a rare gestational trophoblastic tumor and often poses a diagnostic and therapeutic challenge to the involved clinicians. We report a case of epithelioid trophoblastic tumor in a young woman which involved the uterus, parametrium and the right ovary. Misdiagnosis as a choriocarcinoma led to improper treatment and progressive disease. Microscopically it revealed a relatively monotonous population of epithelioid cells arranged in nests with hyaline-like matrix surrounding the tumor cells. Differential diagnosis between placental site trophoblastic tumor and carcinoma was ruled out based on histology and immunohistochemistry. The patient developed lung and brain metastasis after 10 months and is alive with disease 1(1/2) years thereafter and is taking palliative chemotherapy. The patient had beta-HCG level of 85.1 mIU/mL at the time of diagnosis; but just before metastasis, the levels rose. Awareness of the histological features of ETT is essential to avoid misdiagnosis, as it represents a tumor which is primarily treated by surgery rather than with chemotherapy.


Optoacoustic imaging and potential applications of raster-scan optoacoustic mesoscopy in dermatology.
Optoacoustic imaging (OAI) is a hybrid imaging modality that integrates the benefits of optical contrast and ultrasound detection. Raster-scan optoacoustic mesoscopy (RSOM) is an emerging OAI method that provides information about several dermatological conditions' structural, functional, and molecular features. We searched PubMed and Google Scholar databases through September 2021 for contributions relevant to OAI in the English language. This review contains 32 studies and other relevant literature. Several studies indicate that RSOM is helpful in inflammatory skin conditions such as psoriasis and eczema, especially as it allows more accurate quantification of inflammation-related alterations such as changes to the dermal vasculature. In psoriasis, RSOM can provide objective early diagnosis and monitoring of disease activity and treatment efficacy. Multispectral RSOM, a method in which skin is lightened at more than a single wavelength, is beneficial in diagnosing and monitoring hypoxia-associated conditions, such as systemic sclerosis and chronic wounds. OAI techniques can visualize the pathologicvascularization of skin cancers and quantify their oxygenation status which helps differentiate them from normal skin. Also, they can measure the depth of malignant melanoma and detect the metastatic spread of melanoma cells to sentinel lymph nodes. As demonstrated in this contribution, there is a large spectrum of potential applications of OAI imaging, especially RSOM, in diagnosing, treating, and managing skin diseases.


Genetic modifiers of the Drosophila NSF mutant, comatose, include a temperature-sensitive paralytic allele of the calcium channel alpha1-subunit gene, cacophony.
The N-ethylmaleimide-sensitive fusion protein (NSF) has been implicated in vesicle trafficking in perhaps all eukaryotic cells. The Drosophila comatose (comt) gene encodes an NSF homolog, dNSF1. Our previous work with temperature-sensitive (TS) paralytic alleles of comt has revealed a function for dNSF1 at synapses, where it appears to prime synaptic vesicles for neurotransmitter release. To further examine the molecular basis of dNSF1 function and to broaden our analysis of synaptic transmission to other gene products, we have performed a genetic screen for mutations that interact with comt. Here we report the isolation and analysis of four mutations that modify TS paralysis in comt, including two intragenic modifiers (one enhancer and one suppressor) and two extragenic modifiers (both enhancers). The intragenic mutations will contribute to structure-function analysis of dNSF1 and the extragenic mutations identify gene products with related functions in synaptic transmission. Both extragenic enhancers result in TS behavioral phenotypes when separated from comt, and both map to loci not previously identified in screens for TS mutants. One of these mutations is a TS paralytic allele of the calcium channel alpha1-subunit gene, cacophony (cac). Analysis of synaptic function in these mutants alone and in combination will further define the in vivo functions and interactions of specific gene products in synaptic transmission.


Human evolution in Siberia: from frozen bodies to ancient DNA.
The Yakuts contrast strikingly with other populations from Siberia due to their cattle- and horse-breeding economy as well as their Turkic language. On the basis of ethnological and linguistic criteria as well as population genetic studies, it has been assumed that they originated from South Siberian populations. However, many questions regarding the origins of this intriguing population still need to be clarified (e.g. the precise origin of paternal lineages and the admixture rate with indigenous populations). This study attempts to better understand the origins of the Yakuts by performing genetic analyses on 58 mummified frozen bodies dated from the 15th to the 19th century, excavated from Yakutia (Eastern Siberia). High quality data were obtained for the autosomal STRs, Y-chromosomal STRs and SNPs and mtDNA due to exceptional sample preservation. A comparison with the same markers on seven museum specimens excavated 3 to 15 years ago showed significant differences in DNA quantity and quality. Direct access to ancient genetic data from these molecular markers combined with the archaeological evidence, demographical studies and comparisons with 166 contemporary individuals from the same location as the frozen bodies helped us to clarify the microevolution of this intriguing population. We were able to trace the origins of the male lineages to a small group of horse-riders from the Cis-Baïkal area. Furthermore, mtDNA data showed that intermarriages between the first settlers with Evenks women led to the establishment of genetic characteristics during the 15th century that are still observed today.


[High level expression, purification and cytotoxicity of IL-10(18-57)-PE40].
The objective of the experiment is to explore the purification and production of immunotoxin. The chimeric toxin, which is composed of 40 peptides of interleukin 10 (from amino acids 18 to 57) fused to a mutant form of Pseudomonas extoxin (PE) devoid of its native cell recognition domain. Two kinds of prokaryotic expression vector containing the chimeric toxin IL-10(18-57)-PE40 were constructed respectively. After induction of IPTG for 3 hours, IL-10(18-57)-PE40 was expressed highly in cytoplasmic fraction in Rosettablue(DE3), and was directed to periplasmic space as soluble form in E. coli BL21(DE3)pLysS . Western -blotting showed that the expressed protein could react with the specific rabbit sera against LHRH-PE40. With the application of salting out of (NH4)2SO4, hydrophobic interaction chromatography, Cu-affinity chromatography and anion exchange chromatography, the purity of IL-10(18-57)-PE40 was about 96%. The cytotoxicity assay, Cell-ELISA and fluorescent antibody test support the hypothesis that IL-10(18-57) based ligand-mediated cytotoxicity can serve to target cytotoxic agents in vitro.


Conduct of phase I trials in children with cancer.
Future progress in the care of children with cancer requires appropriate evaluations of promising new agents for pediatric indications, beginning with well-conducted phase I trials. This report summarizes current guidelines for the conduct of pediatric phase I trials and represents a consensus between American and European investigators. The primary objective of pediatric phase I trials is to define safe and appropriate doses and schedules of new agents that can subsequently be used in phase II trials to test for activity against specific childhood malignancies. Prioritization of agents for evaluation in children is critical, since many more investigational agents are evaluated in adult patients than can be systematically evaluated in children. Considerations used in prioritizing agents include activity in xenograft models, novel mechanism of action, favorable drug-resistance profile, and activity observed in adult trials of the agent. Distinctive characteristics of pediatric phase I trials, in comparison to adult phase I trials, include the necessity for multiinstitutional participation and their higher starting dose (typically 80% of the adult maximum-tolerated dose [MTD]), both of which reflect the relative unavailability of appropriate patients. The application of uniform eligibility criteria and standard definitions for MTD and dose-limiting toxicity (DLT) help to assure that pediatric phase I trials are safely conducted and reliably identify appropriate doses and schedules of agents for phase II evaluation. Where possible, pediatric phase I trials also define the pharmacokinetic behavior of new agents in children.


A Novel Role for C5a in B-1 Cell Homeostasis.
B-1 cells constitute a unique subpopulation of lymphocytes residing mainly in body cavities like the peritoneal cavity (PerC) but are also found in spleen and bone marrow (BM). As innate-like B cells, they mediate first line immune defense through low-affinity natural IgM (nIgM) antibodies. PerC B-1 cells can egress to the spleen and differentiate into nIgM antibody-secreting plasma cells that recognize conserved exogenous and endogenous cellular structures. Homing to and homeostasis within the PerC are regulated by the chemokine CXCL13 released by PerC macrophages and stroma cells. However, the exact mechanisms underlying the regulation of CXCL13 and B-1 homeostasis are not fully explored. B-1 cells play important roles in the inflammatory response to infection, autoimmunity, ischemia/reperfusion injury, obesity, and atherosclerosis. Remarkably, this list of inflammatory entities has a strong overlap with diseases that are regulated by complement suggesting a link between B-1 cells and the complement system. Interestingly, up to now, no data exist regarding the role of complement in B-1 cell biology. Here, we demonstrate for the first time that C5a regulates B-1 cell steady-state dynamics within the peritoneum, the spleen, and the BM. We found decreased B-1a cell numbers in the peritoneum and the spleen of C5aR1-/- mice associated with increased B1-a and B1-b numbers in the spleen and high serum titers of nIgM antibodies directed against phosphorylcholine and several pneumococcal polysaccharides. Similarly, peritoneal B-1a cells were decreased in the peritoneum and splenic B-1a and B-1b cells were increased in C5aR2-/- mice. The decrease in peritoneal B-1 cell numbers was associated with decreased peritoneal CXCL13 levels in C5aR1-/- and C5aR2-/- mice. In search for mechanisms, we found that combined TLR2 and IL-10 receptor activation in PerC macrophages induced strong CXCL13 production, which was significantly reduced in cells from C5aR1- and C5aR2-deficient mice and after combined C5aR-targeting. Such stimulation also induced marked local C5 production by PerC macrophages and C5a generation. Importantly, peritoneal in vivo administration of C5a increased CXCL13 production. Taken together, our findings suggest that local non-canonical C5 activation in PerC macrophages fuels CXCL13 production as a novel mechanism to control B-1 cell homeostasis.