[Effect of N-nitroso-N-methylurea on DNA and RNA synthesis in cells of liver and spleen tumors in leucosis L-1210].
During the development of leucosis a progressively retarded incorporation of 2-C14-thymidine and C14-uridine into DNA and RNA of a separate tumor cell and an active incorporation of 2-C14-thymidine into DNA of hepatic and splenic cells were observed. Repeated injections of NMU (20 mg/Kg X 5) in tumor-bearing animals result in a twice increase of life terms in mice and suppression of total tumor cells growth. The observed therapeutic effect is related with deep inhibition of DNA and RNA synthesis in leucosis L-1210 cells, while DNA synthesis in hepatic and splenic cells is only insignificantly suppressed (compared with the normal level typical for intact animals), but RNA synthesis in these organs is actually not involved.


New technique for detection of changes in QRS morphology of ECG signals.
With the use of the electrocardiogram (ECG) as a prototype signal, a new technique was devised for detecting signals embedded in noise. Averaged "normal" digitized ECG signals formed a template to which subsequent ECG QRS complexes were compared. The difference between the averaged template signals and subsequent normal beats was white noise, whereas the difference between the template and ectopic beats consisted of nonrandom signal variation. The template to new signal comparison for the zero-, first-, second-, and third-order differences utilized an approximate F test. Accurate detection of abnormal signals associated with high- and low-frequency noise is accomplished with this method, and the practical clinical utility of the method is under study.


Intestinal obstruction caused by omphalomesenteric duct remnant: usefulness of laparoscopy.
The anomalies related to omphalomesenteric duct remnant constitute an uncommon cause of intestinal obstruction, of which Meckel"s diverticulum and its variants represent the most important clinical presentation. In most cases they are asymptomatic and usually affect young patients. When symptomatic, they usually present episodes of gastrointestinal bleeding or acute abdomen syndromes caused by strangulation of intestinal loops as a result of fibrous intraabdominal remnants or inflammation produced by the diverticulum. In most cases, the unexpected presence of these alterations makes intraoperative diagnosis necessary. Treatment is surgical and consists in exeresis of the diverticulum or the fibrous band causing the clinical picture. We report two cases of persistence of the vitelline duct resolved by laparoscopic approach.


High incidence of oesophageal and gastric cancer in Kashmir in a population with special personal and dietary habits.
Over a three year period (1 July 1986 to 30 June 1989) all newly diagnosed and histologically proved cases of oesophageal and gastric cancer were recorded prospectively. Some 1515 cases of oesophageal cancer (1050 men and 465 women) and 966 cases of gastric cancer (789 men and 177 women) were registered. Seven patients had simultaneous oesophageal and gastric cancer. Age standardised incidence rates for oesophageal cancer were: men 43.6/100,000 per year; women 27.9/100,000 per year. The rates for gastric cancer were: men 36.7/100,000 per year, women 9.9/100,000 per annum. These figures were three to six times higher than those recorded by cancer registries in Banglore, Madras, and Bombay. The incidence rates for oesophageal and gastric cancer in Islamabad (southern district of Kashmir) were 4.1 to 5.4 times higher in men and 1.5 to 2.0 times higher in women than those for Kupwara (northern district of Kashmir). The incidence rates for oesophageal and gastric cancer in Muslims, Hindus, and Sikhs were different. The epidemiology of oesophageal cancer in Kashmir was similar to that found in the 'Asian oesophageal cancer belt'. At the same time Kashmir also had an unprecedented high incidence of gastric cancer. Kashmiries have special personal and dietary habits. Further studies are needed to define the relation between these habits and the occurrence of oesophageal and gastric cancer.


Dexrazoxane in combination with anthracyclines lead to a synergistic cytotoxic response in acute myelogenous leukemia cell lines.
In an attempt to improve current therapeutic strategies for acute myelogenous leukemia (AML), we studied the effects of a commercially available drug, dexrazoxane (DEX), which protects against anthracycline-induced cardiotoxicity. The rationale was that DEX would permit higher doses of cardiotoxic drugs to be given. The drug itself may have therapeutic potential as well. Finally, there are concerns that the drug may, as a protective agent, diminish the effectiveness of various chemotherapeutics. To help resolve the question about potential drug antagonism, we undertook a series of in vitro analyses of DEX and various combinations with anthracyclines and other agents. Colony-forming assays were used to evaluate stem-cell renewal of myeloid cells in vitro, and median-effect analysis was used to evaluate antagonism, synergism, and additivity. The anthracyclines doxorubicin, daunorubicin, and idarubicin were individually combined with DEX to study in vitro effects in leukemic myeloid cell lines. In the hope, we could extend the findings to non-anthracyclines, etoposide and cytosine arabinoside were also evaluated in combination with DEX using the same in vitro model and method. We found that the effects of DEX in combination with any of the anthracyclines were schedule dependent. The antitumor effect was greater for each combination than for any anthracycline alone except when DEX was administered 24h before doxorubicin or daunorubicin. These data were corroborated through median-effect analysis. Etoposide in combination with DEX was synergistic for all combinations and schedules, and the combination of cytosine arabinoside and DEX was effective depending on the schedule used. DEX appears to be a promising drug in the treatment of AML and warrants further clinical study involving novel drug combinations.


Karhunen-Loéve expansion of a set of rotated templates.
In this paper, we propose a novel method for efficiently calculating the eigenvectors of uniformly rotated images of a set of templates. As we show, the images can be optimally approximated by a linear series of eigenvectors which can be calculated without actually decomposing the sample covariance matrix.


Impact of prior therapy on the recovery and frequency of corneal pathogens.
To document the impact of prior antibiotic therapy on the recovery of corneal pathogens. Medical records and laboratory reports of 334 consecutive microbial keratitis patients examined from January to December 2000 were reviewed. Comparisons of pathogens, culture positive rate, recovery time, antibiotic sensitivity profile, delay in presentation, and final visual acuity were analyzed for patients treated before presentation and those who were not. The chi square test was used to determine statistical significance. Of the 334 patients, 56% were exposed to at least one course of topical antimicrobials before culture. Patients on therapy were only slightly more likely to be culture negative (P = 0.317) but significantly more likely to have a delay in pathogen recovery (P = 0.002). Patients given prior antibiotics took significantly longer to heal (P = 0.003). Gram-negative organisms (47.5%) were the most frequent pathogens isolated from all culture-positive patients, followed by gram positives (28.7%), fungi (15.8%), and parasites (2%). An increase and significant difference in the frequency of fungi (P = 0.000) and acanthamoeba was reserved for the pretreated group. Gram negative organisms were more often isolated from patients who had not been pretreated (P = 0.002). Pretreated patients were more like to have a pathogen resistant to 1 or more of the commonly prescribed ocular antibiotics (P = 0.02). There is a delay in starting microbiologic-guided antibiotic treatment in patients who have received empiric therapy. Nonbacterial corneal pathogens may be associated more frequently with patients on prior therapy.


Fetal ovarian cysts. Our clinical experience over 16 cases and review of the literature.
Fetal ovarian cysts are intra-abdominal structures frequently diagnosed prenatally, tending to present as isolated unilateral lesions in normal fetuses in the third trimester. These cysts may present with complications and their diameter and echogenicity are the main criteria for establishing their prognosis. Spontaneous regression of fetal ovarian cysts is very usual. In the present study, we present our clinical experience on fetal ovarian cyst surveillance and treatment, as well as a review of the literature in the same field. In this study, we reviewed pre- and postnatal medical records and ultrasonography of 16 fetuses that were diagnosed with ovarian cysts, in Obstetrics Department of University Hospital of Alexandroupolis, between January 2000 and April 2010. We have also reviewed the available literature about fetal ovarian cysts. In a total of 16 cases, postnatal surgery was performed in one infant due to ovarian cyst torsion. In the remaining 15 cases, cysts regressed completely in two fetuses during pregnancy and all the rest of the cysts, including four complex ones, resolved spontaneously after birth. When fetal ovarian cysts are detected, they should be followed up by serial ultrasonographic examinations. The majority of them will regress spontaneously in a period of 12 months after birth, independent of their sonographic findings. Only symptomatic cysts or cysts with a diameter >5 cm, which do not regress or enlarge, should be treated.


[What do intravenous drug users think about hepatitis?].
Intravenous drug users have a high frequency of infectious diseases related to their needle usage, and a higher risk of death. The purpose of this study was to illustrate how intravenous drug users see their health, diseases, and related risks. This article focuses on results related to hepatitis. Seven informants were recruited from a low-threshold civic health centre. The sole criterion for joining the study was previous or current intravenous drug use. Semi-structured interviews were recorded, and the data edited and analysed with a qualitative approach. The informants had experience with and were generally well informed about hepatitis, though some of their perceptions were in disagreement with existing medical knowledge. Self-treatment with intravenous hydrogen peroxide was described as a cure for hepatitis. Despite their accurate knowledge about the risks of infection, the informants still described various methods of sharing their needles and syringes. Their reasons for sharing could be limited access to clean equipment, and an inability to cope. Knowledge alone is inadequate in order to reduce risk behaviour, and a variety of methods, such as access to clean drug equipment, should remain the focus of preventive efforts.


Modification of hippocampal synaptic proteins by nitric oxide-stimulated ADP ribosylation.
Nitric oxide has been shown to be an important neuronal signaling molecule that participates in both behavioral and synaptic plasticity. To better understand the potential mechanisms by which NO regulates synaptic function, the ability of NO to stimulate the modification of synaptic proteins by ADP ribosylation was examined. Two NO donors, sodium nitroprusside and 3-morpholinosydnonimine, stimulated the ADP ribosylation of proteins at apparent molecular masses of 42, 48, 51, 54, and 74 kD in hippocampal synaptosomes. This stimulation was likely owing to the production of NO by the donors; ADP ribosylation was not stimulated by non-NO decomposition products of sodium nitroprusside, and quenching of superoxide anion did not inhibit Sin-1-induced ADP ribosylation. Experiments using NAD+ that was radiolabeled on the nicotinamide moiety demonstrated that the modification of proteins of molecular masses of 30, 33, and 38 kD are not true ADP ribosylation, whereas labeling of the 42-, 48-, 51-, 54-, and 74-kD proteins likely represent ADP ribosylation. Some of the substrates were brain specific (54 and 74 kD), whereas others (42 and 51 kD) were present in multiple nonbrain tissues.


Ankylosing spondylitis with selective IgA deficiency and a circulating anticoagulant.
A patient with ankylosing spondylitis was found to have selective IgA deficiency and a non-heparin, immediate-acting antithrombin (antithrombin V). T cells were decreased, and serum IgG was increased. In vitro synthesis of IgG by peripheral blood lymphocytes was very high. This association of ankylosing spondylitis with the T cell and protein abnormalities is probably fortuitous but does demonstrate that severe spondylitis may evolve in the absence of IgA.


An oroxylin A-loaded aggregation-induced emission active polymeric system greatly increased the antitumor efficacy against squamous cell carcinoma.
Squamous cell carcinoma (SCC) is a usually responds poorly to treatment suffers from poor therapeutic benefits while oroxylin A (OA) is a promising flavonoid with high anticancer efficacy against various cancer types. Here in our study, in order to reveal the potential of OA based drug delivery systems (DDSs) in the treatment of SCC, we firstly revealed that OA had a certain pharmacodynamic effect on skin SCC (A431 cells). Afterwards, OA was loaded into a newly synthesized aggregation-induced emission (AIE)-active polymer to construct OA-loaded PDots for the first time. Our results revealed that OA-loaded PDots showed preferable drug loading and enhanced stability. Moreover, the DDS was also capable of self-illumination in the aggregate state to reveal the uptake profile. Most importantly, the DDS showed much more elevated anticancer benefits than free OA in vitro and advanced tumor targetability in vivo, suggesting that it might be a promising system against SCC.


The Care of "Small Things": Aging and Dignity in Rwanda.
In Rwanda, disruptions to family and social life as a result of the 1994 genocide, and the economic transformations in its aftermath, have complicated the fabric of elder care across the country. In this article, I focus on how elderly Rwandans are reconfiguring their care networks - many of which were destroyed during the genocide - by acting as caregivers and care receivers for each other on a daily basis. Although emotionally and physically taxing, elderly Rwandans emphasize that the "small things" embedded in the giving and receiving of care are intricately connected to how personal and collective dignity is cultivated.


Vernalization response of Phleum pratense and its relationships to stem lignification and floral transition.
Timothy is a long-day grass species well adapted for cultivation in northern latitudes. It produces elongating tillers not only in spring growth but also later in summer. As the quantity and quality of harvested biomass is dictated by canopy architecture and the proportion of stem-forming flowering tillers, the regulation of flowering is of great interest in forage grass production. Canopy architecture, stem morphology and freezing tolerance of vernalized timothy were investigated in greenhouse and field experiments. The molecular control of development was examined by analysing the relationship between apex development and expression of timothy homologues of the floral inducer VRN1 and repressor VRN2. True stem formation and lignification of the sclerenchyma ring occur in both vernalized and regrowing stems irrespective of the developmental stage of the apex. The stems had, however, divergent morphology. Vernalization enhanced flowering, and the expression of the VRN1 homologue was elevated when the apex had passed into the reproductive stage. High VRN1 homologue expression was not associated with reduction in freezing tolerance and the expression coincided with increased levels of the floral repressor VRN2 homologue. Field experiments supported the observed linkage between the upregulation of the VRN1 homologue and the transition to the reproductive stage in vernalized tillers. The upregulation of putative VRN1 or VRN2 genes was restricted to vernalized tillers in the spring yield and, thus, not detected in non-vernalized tillers of the second yield; so-called regrowth. The formation of a lignified sclerenchyma ring that efficiently reduces the digestibility of the stem was not related to apex development but rather to a requirement for mechanical support. The observed good freezing tolerance of reproductive timothy tillers could be one important adaptation mechanism ensuring high yields in northern conditions. Both VRN1 and VRN2 homologues required a vernalization signal for expression so the development of yield-forming tillers in regrowth was regulated independently of the studied genes.


Intravenous immunoglobulin response in new-onset refractory status epilepticus (NORSE) COVID-19 adult patients.
Neurological manifestations may be common in COVID-19 patients. They may include several syndromes, such as a suggested autoimmune abnormal response, which may result in encephalitis and new-onset refractory status epilepticus (NORSE). Quickly recognizing such cases and starting the most appropriate therapy is mandatory due to the related rapid worsening and bad outcomes. This case series describes two adult patients admitted to the university hospital and positive to novel coronavirus 2019 (SARS-CoV-2) infection who developed drug-resistant status epilepticus. Both patients underwent early electroencephalography (EEG) assessment, which showed a pathological EEG pattern characterized by general slowing, rhythmic activity and continuous epileptic paroxysmal activity. A suspected autoimmune etiology, potentially triggered by SARS-CoV-2 infection, encouraged a rapid work-up for a possible autoimmune encephalitis diagnosis. Therapeutic approach included the administration of 0.4 g/kg intravenous immunoglobulin, which resulted in a complete resolution of seizures after 5 and after 10 days, respectively, without adverse effects and followed by a normalization of the EEG patterns.


[Antiviral and antiproliferative activity of human interferons and their induction of 2',5'-oligoadenylate synthetase].
Native preparations of alpha, beta and gamma-interferons as well as recombinant beta-interferon and purified leukocyte alpha-interferon and purified leukocyte alpha-interferon exert antiviral and antiproliferative activity in CaOv cells. Native interferon preparations were shown to be more antiproliferative than purified interferons per unit of antiviral activity (with EMC as well as with less susceptible VSV used as test viruses). It was shown that level of 2'5' oligoadenylatesynthetase activity induction in general correlates with antiproliferative and pronounced antiviral activity of interferons, besides that, the earlier (by 11 hours) induction of the enzyme activity by beta-interferon correlates with more rapid expression of antiproliferative effects by this interferon in comparison with that of alpha-interferon, the latter inducing the peak of enzyme activity by 24 hours.


Use of the Galleria mellonella caterpillar as a model host to study the role of the type III secretion system in Pseudomonas aeruginosa pathogenesis.
Nonvertebrate model hosts represent valuable tools for the study of host-pathogen interactions because they facilitate the identification of bacterial virulence factors and allow the discovery of novel components involved in host innate immune responses. In this report, we determined that the greater wax moth caterpillar Galleria mellonella is a convenient nonmammalian model host for study of the role of the type III secretion system (TTSS) in Pseudomonas aeruginosa pathogenesis. Based on the observation that a mutation in the TTSS pscD gene of P. aeruginosa strain PA14 resulted in a highly attenuated virulence phenotype in G. mellonella, we examined the roles of the four known effector proteins of P. aeruginosa (ExoS, ExoT, ExoU, and ExoY) in wax moth killing. We determined that in P. aeruginosa strain PA14, only ExoT and ExoU play a significant role in G. mellonella killing. Strain PA14 lacks the coding sequence for the ExoS effector protein and does not seem to express ExoY. Moreover, using Delta exoU Delta exoY, Delta exoT Delta exoY, and Delta exoT Delta exoU double mutants, we determined that individual translocation of either ExoT or ExoU is sufficient to obtain nearly wild-type levels of G. mellonella killing. On the other hand, data obtained with a Delta exoT Delta exoU Delta exoY triple mutant and a Delta pscD mutant suggested that additional, as-yet-unidentified P. aeruginosa components of type III secretion are involved in virulence in G. mellonella. A high level of correlation between the results obtained in the G. mellonella model and the results of cytopathology assays performed with a mammalian tissue culture system validated the use of G. mellonella for the study of the P. aeruginosa TTSS.


Hsp90 Is Essential under Heat Stress in the Bacterium Shewanella oneidensis.
The Hsp90 chaperone is essential in eukaryotes and activates a large array of client proteins. In contrast, its role is still elusive in bacteria, and only a few Hsp90 bacterial clients are known. Here, we found that Hsp90 is essential in the model bacterium Shewanella oneidensis under heat stress. A genetic screen for Hsp90 client proteins identified TilS, an essential protein involved in tRNA maturation. Overexpression of TilS rescued the growth defect of the hsp90 deletion strain under heat stress. In vivo, the activity and the amount of TilS were significantly reduced in the absence of Hsp90 at high temperature. Furthermore, we showed that Hsp90 interacts with TilS, and Hsp90 prevents TilS aggregation in vitro at high temperature. Together, our results indicate that TilS is a client of Hsp90 in S. oneidensis. Therefore, our study links the essentiality of bacterial Hsp90 at high temperature with the identification of a client.


Study of Staphylococcus aureus adhesion on a novel nanostructured surface by chemiluminometry.
In recent years the progress in the field of nanotechnologies has offered new possibilities to control the superficial features of implant materials down to a nanoscale level. Several studies have therefore tried to explore the effects of nanostructured biomaterial surfaces on the behavior of eukaryotic cells. However, nanotopography could exert an influence also on the behavior of prokaryotic cells, with relevant implications concerning the susceptibility of implant surfaces to infection. Aim of this study was to examine the behavior of Staphylococcus aureus on polyethylene terephthalate (PET) surfaces either cylindrically nanostructured (PET-N) or flat ion-etched (PET-F), and on tissue culture-grade polystyrene (PS). Microbial adherence was assessed by chemiluminometry under 4 different conditions: (a) bacteria suspended in MEM medium, (b) bacteria in MEM supplemented with 10% fetal bovine serum (FBS), (c) test surfaces preconditioned in FBS, and (d) post-exposure of colonised surfaces to serum-supplemented MEM. Under all circumstances, PET-F and PET-N specimens showed identical bacterial adhesion properties. In the absence of serum, all 3 test materials showed a very high adhesivity to microbial cells and both PET surfaces exhibited greater adhesion than PS. On the contrary, the presence of 10% serum in solution significantly affected cell behavior: the number of microbial cells on all surfaces was drastically reduced, and the adhesion properties of PET surfaces with respect to PS were reversed, with PET being less adhesive. Overall, the specific cylindrical nanostructures created on PET did not significantly influence microbial behavior. Ongoing studies are verifying whether other nanotopographies with different geometry could have more substantial effects.


Four Novel Motion Paradigms Based on Steady-State Motion Visual Evoked Potential.
The purpose of this paper was to study the applicability of paradigms with motion forms for use in a brain-computer interface (BCI). We examined the performances of different paradigms and evaluated the stimulus effects. We designed four novel stimulus paradigms based on basic motion modes: swing, rotation, spiral, and radial contraction-expansion. Canonical correlation analysis (CCA) was used to analyze the accuracy. Additionally, we optimized CCA template signal harmonic combinations for the different motion paradigms. The spiral motion paradigm exhibited the highest average information transfer rate (ITR) and recognition accuracy (41.24 bit/min-1/95.33%), and the average ITRs and recognition accuracies were lowest for the rotation motion paradigm (31.89 bit/min-1 /80.89%) and the radial contraction-expansion motion paradigm (32.62 bit/min-1 /80.72%) because they include fewer harmonic components. Any stimulus paradigms with periodic motion can induce steady-state motion visual evoked potentials (SSMVEPs), but the SSMVEP harmonic components induced by different motion modes differed significantly. The spiral motion paradigm was more suitable for BCI applications. This study is an important extension to the existing SSMVEP-based BCI literature, and provides new insight to enable future design of the BCI paradigms.


Differences in the use and appreciation of a web-based or printed computer-tailored physical activity intervention for people aged over 50 years.
This study provides insight into the use and appreciation of a tailored physical activity intervention for people aged over 50 years in different intervention conditions (i.e. printed versus web-based and basic versus environmental). Participants (within a clustered randomized controlled trial) received printed or web-based-tailored advice three times within 4 months. Half of the participants also received environmental information. Differences in use and appreciation between both delivery modes and between the basic and environmental condition (similar delivery mode) were assessed at 3 (N = 935) and 6 (N = 649) months after baseline using analyses of variance and chi-square test. The use of the printed intervention (i.e. 92.7-98.2% read, 70.1-76.5% kept and 39.9-56.8% discussed) was significantly higher and printed intervention components were better appreciated than web-based intervention (scores, respectively, 6.06-6.91 versus 5.05-6.11 on a scale of 1-10). In-depth appreciation (e.g. reliability, perceived individualization) was average to high, without differences between intervention conditions. Additional environmental information did not increase appreciation; however, environmental intervention components were more used compared with basic intervention components. Integration of environmental components can stimulate active use of the intervention. To increase the public impact and prevent dropout by participants of web-based physical activity interventions, design modifications are needed for ease of use and improved appreciation.


Host-guest complexes of conformationally flexible C-hexyl-2-bromoresorcinarene and aromatic N-oxides: solid-state, solution and computational studies.
Host-guest complexes of C-hexyl-2-bromoresorcinarene (BrC6) with twelve potential aromatic N-oxide guests were studied using single crystal X-ray diffraction analysis and 1H NMR spectroscopy. In the solid state, of the nine obtained X-ray crystal structures, eight were consistent with the formation of BrC6-N-oxide endo complexes. The lone exception was from the association between 4-phenylpyridine N-oxide and BrC6, in that case the host forms a self-inclusion complex. BrC6, as opposed to more rigid previously studied C-ethyl-2-bromoresorcinarene and C-propyl-2-bromoresorcinarene, undergoes remarkable cavity conformational changes to host different N-oxide guests through C-H···π(host) interactions. In solution phase CD3OD/CDCl3 (1:1 v/v), all twelve N-oxide guests form endo complexes according to 1H NMR; however, in more polar CD3OD/DMSO-d6 (9:1 v/v), only three N-oxides with electron-donating groups form solution-phase endo complexes with BrC6. In solid-state studies, 3-methylpyridine N-oxide+BrC6 crystallises with both the upper- and lower-rim BrC6 cavities occupied by N-oxide guests. Computational DFT-based studies support that lower-rim long hexyl chains provide the additional stability required for this ditopic behaviour. The lower-rim cavity, far from being a neutral hydrophobic environment, is a highly polarizable electrostatically positive surface, aiding in the binding of polar guests such as N-oxides.


Spawning migration of American eel Anguilla rostrata from pristine (1843-1872) to contemporary (1963-1990) periods in the St Lawrence Estuary, Canada.
Daily American eel Anguilla rostrata catches and their dates of passage (starting, median and ending dates) were compared between pristine (1843-1872) and contemporary periods (1963-1990) to determine any changes and see whether these were related to environmental variations and water discharge regulation. Timing and duration of A. rostrata migration patterns differed significantly between the two periods. In the contemporary period, migrating A. rostrata were intercepted significantly earlier than in pristine times (18 days earlier on average), and ended at the same average period. Early A. rostrata migration was also significantly related to high spring flow and secondarily to high spring temperature, while migration ended later when high temperature or low water level occurred during the autumn period. A recent slight increase in the water temperature of the St Lawrence River could partially explain the earlier A. rostrata migration observed during the contemporary period. In return, the effect of high spring flow should have been more contrasted if the river discharge would have not been regulated. Recent A. rostrata production now being mainly restricted to the lower part of St Lawrence River mainstream, resulting shorter travelling distance to the estuary may explain why migrating progress was earlier during the contemporary period.


Dynamin- and Rab5-dependent endocytosis is required to prevent Drosophila photoreceptor degeneration.
In Drosophila photoreceptors, Rhodopsin 1 (ninaE, Rh1) is required for proper morphogenesis and maintenance of the apical light-gathering organelle, the rhabdomere. It has been proposed that Rh1, coupled to the Rho GTPases Rac1 and Cdc42, promotes the morphogenesis of a sub-rhabdomeric F-actin meshwork or rhabdomere terminal web (RTW). The RTW provides mechanical support to the apical microvilli and is likely to guide Rab11-dependent delivery of Rh1-rich membrane to the rhabdomere from the trans Golgi network. However, the nature and function of the molecular pathway involved in RTW morphogenesis remains incomplete. Here, we show that Rh1 function in promoting RTW morphogenesis is light-independent and is conserved throughout evolution. This Rh1 function does not require G(q)α(e), which is required for phototransduction. Finally, we show that interfering with Dynamin- and Rab5-dependent endocytosis leads to a phenotype that is undistinguishable from that of the ninaE-null mutant. Importantly, the corresponding endocytic activity is essential to prevent early onset of rhabdomere degeneration. In conclusion, we propose that Rh1 function in promoting RTW morphogenesis is not only needed to sustain apical membrane delivery but is also required for proper rhabdomeric membrane endocytosis and turnover.


Does managed care affect the diffusion of psychotropic medications?
Newer technologies to treat many mental illnesses have shown substantial heterogeneity in diffusion rates across states. In this paper, I investigate whether variation in the level of managed care penetration is associated with changes in state-level diffusion of three newer classes of psychotropic medications in fee-for-service Medicaid programs from 1991 to 2005. Three different types of managed care programs are examined: capitated managed care, any type of managed care and behavioral health carve-outs. A fourth-order polynomial fixed effect regression model is used to model the diffusion path of newer antidepressant and antipsychotic medications controlling for time-varying state characteristics. Substantial differences are found in the diffusion paths by the degree of managed care use in each state Medicaid program. The largest effect is seen through spillover effects of capitated managed care programs; states with greater capitated managed care have greater initial shares of newer psychotropic medications. The influence of carve-outs and of all types of managed care combined on the diffusion path was modest.


2,5-Disubstituted tetrahydrofurans as selective serotonin re-uptake inhibitors.
Enhancement of 5-hydroxytryptamine (5-HT, serotonin) neurotransmission is a viable means of treating depression. On the basis of this observation, agents that inhibit re-uptake of 5-HT were prepared based on (-)-cocaine and aryltropanes as lead compounds because they are reasonably potent 5-HT re-uptake inhibitors. Molecular dissection of an aryltropane provided a series of 5- and 6-membered ring compounds. From among this library of compounds a series of disubstituted tetrahydrofurans bearing 2-alkyl aryl and 5-alkyl amino groups were identified as having highly potent and selective 5-HT re-uptake inhibition. The compounds were evaluated for their ability to compete with radiolabeled RTI-55 binding and to inhibit re-uptake of neurotransmitters at the human dopamine, serotonin and norepinephrine transporters. Based on potency (e.g., K(i)=800 pM) and significant functional selectivity (e.g., IC(50) ratios for human dopamine:serotonin or norepinephrine:serotonin, >or=1397) highly potent and selective serotonin re-uptake inhibitors were identified. Optimal features playing a dominant role in binding affinity and re-uptake inhibition included lipophilic substitution on the aromatic moiety, trans relative stereochemistry of the 2,5-disubstituted tetrahydrofuran ring, and a total of four or five methylene groups between the alkyl amine and the alkyl aryl moiety and the tetrahydrofuran group. A number of the most potent serotonin re-uptake inhibitors were tested in Balb/c mice in the forced-swim test (FST), a behavioral test used to measure the effects of antidepressant agents. Acute administration of 32c (10mg/kg), or 32d (10mg/kg) ip tended to decrease the duration of mouse immobility in the FST although the effect was not statistically significant.


Description of a new disposable uterine sampler (the Accurette) for endometrial cytology and histology.
A disposable, collapsible curette made from a special plastic material, devised by the author for obtaining cytological and histological samples of the uterine endometrium, is described. A simple syringe for vacuum aspiration is incorporated with the instrument. Its manifold advantages include simplicity of design and ease of application, making little demand on the operator. No additional apparatus is required for its use in outpatient clinics or as an office procedure.


Furfural production by 'acidic steam stripping' of lignocellulose.
Furfural and acetic acid are produced with approximately 60 and 90 mol % yield, respectively, upon stripping bagasse with a gaseous stream of HCl/steam and condensing the effluent to water/furfural/acetic acid. The reaction kinetics is 1(st) order in furfural and 0.5(th) order in HCl. A process concept with full recycling of the reaction effluents is proposed to reduce the energy demand to <10 tonsteam tonfurfural (-1) and facilitate the product recovery through a simple liquid/liquid separation of the condensate into a water-rich and a furfural-rich phase.


Norethisterone, a major ingredient of contraceptive pills, is a suicide inhibitor of estrogen biosynthesis.
Norethisterone (17 alpha-ethynyl-19-nortestosterone) is an effective irreversible inhibitor of estrogen synthetase (aromatase), the enzyme responsible for the conversion of androgens to estrogens, even at a 2 X 10(-6) molar concentration. This irreversible inactivation, which is directed toward the active site of aromatase and requires the cofactor-reduced nicotinamide adenine dinucleotide phosphate, is both time- and concentration-dependent. Ethisterone (17 alpha-ethynyltestosterone), in contrast, is not a suicide inhibitor of aromatase even at concentrations of 10(-4) molar.


Standardized grey scale ultrasonography in Graves' disease: correlation to autoimmune activity.
Graves' disease leads to thyroid enlargement and to reduction of tissue echogenicity. Our purpose was to correlate grey scale ultrasonography of the thyroid gland with clinical and laboratory findings in patients with Graves' disease. Fifty-three patients with Graves'disease were included in our study, 100 euthyroid volunteers served as control group. Free thyroxine (FT(4)), TSH and TRAb (TSH receptor antibodies) values were measured and correlated with sonographic echogenicity of the thyroid gland. All patients and control persons underwent ultrasonographical histogram analyses under standardized conditions. Mean densities of the thyroid tissues were determined in grey scales (GWE). Compared with controls with homogeneous thyroid lobes of normal size (25.6 +/- 2.0GWE, mean +/- S.D.) echogenicity in patients with Graves' disease was significantly lower (21.3 +/- 3. 3GWE, mean +/- S.D., P < 0.0001). Among the patients with Graves' disease significant differences of thyroid echo levels were revealed for patients with suppressed (20.4 +/- 3.1 GWE, mean +/- S.D., n=34) and normalized TSH values (22.5 +/- 3.6GWE, mean +/- S.D., n=19, P < 0.02). Significantly lower echogenicities were also measured in cases of persistent elevated TRAb levels (19.9 +/- 2.9GWE, mean +/- S.D., n=31) in comparison with normal TRAb levels (22.9 +/- 3.5 GWE, mean +/- S.D., n=22, P < 0.0015). No correlation could be verified between echogenicity and either still elevated or already normalized FT(4) values or the thyroid volume. In coincidence of hyperthyroidism and Graves' ophthalmopathy (19.7 +/- 3.5GWE, mean +/- S.D., n=23) significantly lower echogenicity was measured than in the absence of ophthalmological symptoms (22.3 +/- 3.3GWE, mean +/- S.D., n=30, P < 0.016). Patients needing active antithyroid drug treatment revealed significantly lower thyroid echogenicity (20.3 +/- 3.1 GWE, mean +/- S.D., n=40) than patients in remission (23.7 +/- 3.4 GWE, mean +/- S.D., n=13, P < 0.001). Statistical evaluation was carried out using Student's t-test. Standardized grey scale histogram analysis allows for supplementary judgements of thyroid function and degree of autoimmune activity in Graves' disease. Whether these values help to estimate the risk of recurrence of hyperthyroidism after withdrawal of antithyroid medication should be evaluated in a prospective study.


Gaussian imaging transformation for the paraxial Debye formulation of the focal region in a low-Fresnel-number optical system
The Debye formulation of focused fields has been systematically used to evaluate, for example, the point-spread function of an optical imaging system. According to this approximation, the focal wave field exhibits some symmetries about the geometrical focus. However, certain discrepancies arise when the Fresnel number, as viewed from focus, is close to unity. In that case, we should use the Kirchhoff formulation to evaluate accurately the three-dimensional amplitude distribution of the field in the focal region. We make some important remarks regarding both diffraction theories. In the end we demonstrate that, in the paraxial regime, given a defocused transverse pattern in the Debye approximation, it is possible to find a similar pattern but magnified and situated at another plane within the Kirchhoff theory. Moreover, we may evaluate this correspondence as the action of a virtual thin lens located at the focal plane and whose focus is situated at the axial point of the aperture plane. As a result, we give a geometrical interpretation of the focal-shift effect and present a brief comment on the problem of the best-focus location.


Electrical stimulation thresholds differ between V1 and V2.
Cortical visual prostheses are designed to treat blindness by restoring visual perceptions through artificial electrical stimulation of the primary visual cortex (V1). Intracortical microelectrodes produce the smallest visual percepts and thus higher resolution vision - like a higher density of pixels on a monitor. However, intracortical microelectrodes must maintain a minimum spacing to preserve tissue integrity. One solution to increase the density of percepts is to implant and stimulate multiple visual areas, such as V1 and V2, although the properties of microstimulation in V2 remain largely unexplored. We provide a direct comparison of V1 and V2 microstimulation in two common marmoset monkeys. We find similarities in response trends between V1 and V2 but differences in threshold, neural activity duration, and spread of activity at the threshold current. This has implications for using multi-area stimulation to increase the resolution of cortical visual prostheses.


Structural requirements of lipid A species in activation of clotting enzymes from the horseshoe crab, and the human complement cascade.
The structure/activity relationship of lipid A, a bioactive center of endotoxic lipopolysaccharides, in the activation of the clotting enzyme cascade of a horseshoe crab amoebocyte lysate (Limulus activity) and the complement system in human serum, was examined using synthetic lipids A and related compounds. Regarding Limulus activity, a newly developed colorimetric method, which utilizes a mixture of recombined clotting factors and a chromogenic substance, was much more sensitive for detecting changes in the chemical structure of test compounds than the conventional gelation method using the amoebocyte whole lysate. (beta 1-6)-D-Glucosamine disaccharide bisphosphates, which had neither 3-hydroxyacyl nor 3-acyloxyacyl groups, and acylglucosamine phosphates, which in structure correspond or are analogous to the non-reducing or reducing moieties of lipids A and biosynthetic disaccharide lipid A precursors showed only negligible activity in the colorimetric tests, but they exhibited a distinct though much weaker gelation activity than the parent disaccharide molecules. The assay results obtained by the colorimetric Limulus test correlate better with the pyrogenicity of the test synthetic compounds than those given by the gelation method, although the dependence of pyrogenicity on chemical structure is greater. The presence of 3-hydroxyacyl groups on the bisphosphorylated (beta 1-6)-D-glucosamine disaccharide backbone is the prerequisite for effective activation of the clotting enzyme cascade of horseshoe crab amoebocyte lysate, while the presence of an adequate number (one or two) of 3-acyloxyacyl groups on the disaccharide bisphosphate backbone is needed for full pyrogenicity. Complement activation, on the other hand, showed structural requirements quite different from those for the colorimetric Limulus activity and the pyrogenicity. The disaccharide compounds that had only non-hydroxylated acyl groups, acylated glucosamine phosphates that had the structure of the non-reducing portion of lipids A and biosynthetic disaccharide precursors, which were scarcely active in the colorimetric Limulus test, caused complement activation comparable to or sometimes stronger than that of the parent disaccharide molecules. Acylglucosamine phosphates, corresponding in structure to the reducing moiety of disaccharide compounds, however, showed little activity.


Regulation of gene expression by the small GTPase Rho through the ERK6 (p38 gamma) MAP kinase pathway.
Small GTP-binding proteins of the Rho-family, Rho, Rac, and Cdc42, have been traditionally linked to the regulation of the cellular actin-based cytoskeleton. Rac and Cdc42 can also control the activity of JNK, thus acting in a molecular pathway transmitting extracellular signals to the nucleus. Interestingly, Rho can also regulate gene expression, albeit by a not fully understood mechanism. Here, we found that activated RhoA can stimulate c-jun expression and the activity of the c-jun promoter. As the complexity of the signaling pathways controlling the expression of c-jun has begun to be unraveled, this finding provided a unique opportunity to elucidate the biochemical routes whereby RhoA regulates nuclear events. We found that RhoA can initiate a linear kinase cascade leading to the activation of ERK6 (p38 gamma), a recently identified member of the p38 family of MAPKs. Furthermore, we present evidence that RhoA, PKN, MKK3/MKK6, and ERK6 (p38 gamma) are components of a novel signal transduction pathway involved in the regulation of gene expression and cellular transformation.


Duplication Xp22.2 and pseudoisodicentric Yq detected by FISH and PCR in a sterile male.
A chromosome mosaicism with two cell lines was diagnosed in a sterile man. One cell line had a 45, -Y, dup (X) (p22.2) karyotype and accounted for 83% of lymphocytes analyzed. Fluorescence in situ hybridization (FISH) analysis with specific X and Y probes excluded a translocation between the short arms of the X and Y chromosomes and showed that Xp duplication involved a region containing the DXS85 locus, distal to the ZFX and DSS sites. The other cell line consisted of a diploid karyotype with a rearranged Y chromosome, which was shown to be a pseudoisodicentric Yq by FISH. Moreover, FISH with a specific probe for the AZF locus and polymerase chain reaction using Yq SY108 and SY121 primers showed no signals for this region, possibly accounting for the azoospermia in this patient.


Contrasting growth responses of dominant peatland plants to warming and vegetation composition.
There is growing recognition that changes in vegetation composition can strongly influence peatland carbon cycling, with potential feedbacks to future climate. Nevertheless, despite accelerated climate and vegetation change in this ecosystem, the growth responses of peatland plant species to combined warming and vegetation change are unknown. Here, we used a field warming and vegetation removal experiment to test the hypothesis that dominant species from the three plant functional types present (dwarf-shrubs: Calluna vulgaris; graminoids: Eriophorum vaginatum; bryophytes: Sphagnum capillifolium) contrast in their growth responses to warming and the presence or absence of other plant functional types. Warming was accomplished using open top chambers, which raised air temperature by approximately 0.35 °C, and we measured air and soil microclimate as potential mechanisms through which both experimental factors could influence growth. We found that only Calluna growth increased with experimental warming (by 20%), whereas the presence of dwarf-shrubs and bryophytes increased growth of Sphagnum (46%) and Eriophorum (20%), respectively. Sphagnum growth was also negatively related to soil temperature, which was lower when dwarf-shrubs were present. Dwarf-shrubs may therefore promote Sphagnum growth by cooling the peat surface. Conversely, the effect of bryophyte presence on Eriophorum growth was not related to any change in microclimate, suggesting other factors play a role. In conclusion, our findings reveal contrasting abiotic and biotic controls over dominant peatland plant growth, suggesting that community composition and carbon cycling could be modified by simultaneous climate and vegetation change.


Operando Laboratory X-Ray Imaging of Silver-Based Gas Diffusion Electrodes during Oxygen Reduction Reaction in Highly Alkaline Media.
Operando laboratory X-ray radiographies were carried out for imaging of two different silver-based gas diffusion electrodes containing an electroconductive Ni mesh structure, one gas diffusion electrode composed of 95 wt.% Ag and 5 wt.% polytetrafluoroethylene and one composed of 97 wt.% Ag and 3 wt.% polytetrafluoroethylene, under different operating parameters. Thereby, correlations of their electrochemical behavior and the transport of the 30 wt.% NaOH electrolyte through the gas diffusion electrodes were revealed. The work was divided into two parts. In the first step, the microstructure of the gas diffusion electrodes was analyzed ex situ by a combination of focused ion beam technology and synchrotron as well as laboratory X-ray tomography and radiography. In the second step, operando laboratory X-ray radiographies were performed during chronoamperometric measurements at different potentials. The combination of the ex situ microstructural analyses and the operando measurements reveals the impact of the microstructure on the electrolyte transport through the gas diffusion electrodes. Hence, an impact of the Ni mesh structure within the gas diffusion electrode on the droplet formation could be shown. Moreover, it could be observed that increasing overpotentials cause increasing electrolyte transport velocities and faster droplet formation due to electrowetting. In general, higher electrolyte transport velocities were found for the gas diffusion electrode with 97 wt.% Ag in contrast to that with 95 wt.% Ag.


Management of epilepsy in resource-limited settings.
Epilepsy is one of the most common and widespread neurological disorders affecting over 65 million people worldwide. Although estimates vary considerably, the annual incidence is considered to be almost 50 per 100,000 and prevalence around 700 per 100,000. It is thought, however, that more people are affected in low- and middle-income countries where resources to improve the care for people with epilepsy are limited. Of all people with epilepsy, around 80% live in resource-limited countries and up to 90% of these patients receive no treatment at all. National epilepsy programs to organize comprehensive care and cover educational, economic, and research aspects are necessary. A referral network will enable local healthcare workers to consult patients with more complex diseases and may ensure routine availability of inexpensive AEDs. Adequately identifying people with epilepsy and delivering cost-effective care in resource-limited countries is an important challenge for epileptologists and healthcare policy makers alike. Here we give an overview of the present situation and review the needs and the efforts currently being made in the field.


Direct Wiring of Liquid Metal on an Ultrasoft Substrate Using a Polyvinyl Alcohol Lift-off Method.
In recent years, wiring and system construction on ultrasoft materials such as biological tissues and hydrogels have been proposed for advanced wearable devices, implantable devices, and soft robotics. Among the soft conductive materials, Ga-based liquid metals (LMs) are both biocompatible and ultrasoft, making them a good match for electrodes on the ultrasoft substrates. However, gels and tissues are softer and less wettable to the LMs than conventional soft substrates such as Ecoflex and polydimethylsiloxane. In this study, we demonstrated the transfer of LM paste composed of Ga-based LM and Ni nanoparticles onto ultrasoft substrates such as biological tissue and gels using sacrificial polyvinyl alcohol (PVA) films. The LM paste pattern fabricated on the PVA film adhered to the ultrasoft substrate along surface irregularities and was transferred without being destroyed by the PVA film before the PVA's dissolution in water. The minimum line width that could be wired was approximately 165 μm. Three-dimensional wiring, such as the helical structure on the gel fiber surface, is also possible. Application of this transfer method to tissues using LM paste wiring allowed the successful stimulation of the vagus nerve in rats. In addition, we succeeded in transferring a temperature measurement system fabricated on a PVA film onto the gel. The connection between the solid-state electrical element and the LM paste was stable and maintained the functionality of the temperature-sensing system. This fundamental study of wiring fabrication and system integration can contribute to the development of advanced electric devices based on ultrasoft substrates.


Structural genomic variation in ischemic stroke.
Technological advances in molecular genetics allow rapid and sensitive identification of genomic copy number variants (CNVs). This, in turn, has sparked interest in the function such variation may play in disease. While a role for copy number mutations as a cause of Mendelian disorders is well established, it is unclear whether CNVs may affect risk for common complex disorders. We sought to investigate whether CNVs may modulate risk for ischemic stroke (IS) and to provide a catalog of CNVs in patients with this disorder by analyzing copy number metrics produced as a part of our previous genome-wide single-nucleotide polymorphism (SNP)-based association study of ischemic stroke in a North American white population. We examined CNVs in 263 patients with ischemic stroke (IS). Each identified CNV was compared with changes identified in 275 neurologically normal controls. Our analysis identified 247 CNVs, corresponding to 187 insertions (76%; 135 heterozygous; 25 homozygous duplications or triplications; 2 heterosomic) and 60 deletions (24%; 40 heterozygous deletions; 3 homozygous deletions; 14 heterosomic deletions). Most alterations (81%) were the same as, or overlapped with, previously reported CNVs. We report here the first genome-wide analysis of CNVs in IS patients. In summary, our study did not detect any common genomic structural variation unequivocally linked to IS, although we cannot exclude that smaller CNVs or CNVs in genomic regions poorly covered by this methodology may confer risk for IS. The application of genome-wide SNP arrays now facilitates the evaluation of structural changes through the entire genome as part of a genome-wide genetic association study.


A simple yet effective AIE-based fluorescent nano-thermometer for temperature mapping in living cells using fluorescence lifetime imaging microscopy.
We designed and synthesized a novel nano-thermometer using aggregation-induced-emission (AIE) dye as the reporter and household butter as the matrix. This temperature nanosensor showed decreased fluorescence intensities (∼2%/°C) and shorter fluorescence lifetimes (∼0.11 ns/°C) upon increasing the environmental temperature in the physiological temperature range. Such fluorescence responses were reversible and independent of the environmental pH and ionic strength. The application of these nano-thermometers in temperature sensing in living cells using fluorescence lifetime imaging microscopy (FLIM) was also demonstrated. To the best of our knowledge, this is the first example of AIE-based nano-thermometer for temperature sensing in living cells. This work also provides us with a simple and low-cost method for rapid fabrication of an effective nanosensor based on AIE mechanism.


13q deletion anatomy and disease progression in patients with chronic lymphocytic leukemia.
Historically, genes targeted by recurrent chromosomal deletions have been identified within the smallest genomic region shared in all patients, the minimally deleted region (MDR). However, deletions this small do not occur in all patients and are a simplification of the impact larger heterogeneous deletions have during carcinogenesis. We use the example of 13q14 deletions in chronic lymphocytic leukemia to show that genes outside MDRs are associated with disease progression. Genomic profiling of 224 patients identified 205 copy number alterations on chromosome 13 in 132 cases. Deletions including DLEU2 were heterogeneous (845 Kb-96.2 Mb) and identified two breakpoint cluster regions within short interspersed nuclear elements proximal to DLEU2 and within long interspersed nuclear elements/L1 repeats distal to GUCY1B2. After defining a deletion class on the basis of size and location, we show that (a) at diagnosis, larger deletions (class II) were associated with a significantly increased risk of disease progression (odds ratio=12.3; P=0.005), (b) in progressive patients, class II deletions were enriched (P=0.02) and (c) this association was independent of IgVH mutational status, ZAP70 expression and ATM/TP53 deletion. Deletion of a 1 Mb gene cluster (48.2-49.2 Mb), including SETDB2, PHF11 and RCBTB1, was significantly associated (P<0.01) with disease progression. Here, we show that the deletion of genes outside MDRs can influence clinical outcome.


Rituximab Use and Hypogammaglobulinemia.
BACKGROUND Rituximab is a genetically engineered chimeric (murine-human) monoclonal antibody (mAb) directed against CD20 antigen on the surface of B cells. Commonly reported adverse effects are chills and fevers, which are usually associated with the first infusion. Recent studies have shown an association between rituximab use and low immunoglobulin (Ig) level due to a reduction in plasma cell precursors, which leads to an increased risk of infections with the use of rituximab. CASE REPORT We present a case of hypogammaglobulinemia associated with rituximab use in a patient with Granulomatosis with Polyangiitis (GPA). A 59-year-old woman presented with shortness of breath. After an extensive workup, she was diagnosed with GPA. She received rituximab for the induction of remission. Laboratory workup, which was done five days after she received the second rituximab dose, showed IgG and IgM levels below the level of normal. One month after her second dose of rituximab, she presented to the Medical Intensive Care Unit as a transfer from an outside facility intubated and sedated due to acute respiratory failure secondary to septic shock with E. coli bacteremia. The patient died on admission despite aggressive management, including the ACLS protocol. CONCLUSIONS Rituximab is an effective medication in the management of ANCA-associated vasculitis. Obtaining an immunoglobulin level at baseline and before each rituximab cycle is of great clinical importance and helps guide physicians in prescribing B cell-targeted therapy.


Effects of acute kidney injury after liver resection on long-term outcomes.
To investigate the effects of acute kidney injury (AKI) after liver resection on the long-term outcome, including mortality and renal dysfunction after hospital discharge. We conducted a historical cohort study of patients who underwent liver resection for hepatocellular carcinoma with sevoflurane anesthesia between January 2004 and October 2011, survived the hospital stay, and were followed for at least 3 years or died within 3 years after hospital discharge. AKI was diagnosed based on the Acute Kidney Injury Network classification within 72 hours postoperatively. In addition to the data obtained during hospitalization, serum creatinine concentration data were collected and the glomerular filtration rate (GFR) was estimated after hospital discharge. AKI patients (63%, P = 0.002) were more likely to reach the threshold of an estimated GFR (eGFR) of 45 ml/min/1.73 m2 within 3 years than non-AKI patients (31%) although there was no significant difference in mortality (33% vs. 29%). Cox proportional hazard regression analysis showed that postoperative AKI was significantly associated with the composite outcome of mortality or an eGFR of 45 ml/min/1.73 m2 (95% CI of hazard ratio, 1.05-2.96, P = 0.033), but not with mortality (P = 0.699), the composite outcome of mortality or an eGFR of 60 ml/min/1.73 m2 (P =0.347). After liver resection, AKI patients may be at higher risk of mortality or moderate renal dysfunction within 3 years. These findings suggest that even after discharge from the hospital, patients who suffered AKI after liver resection may need to be followed-up regarding renal function in the long term.


Cortical myoclonus and epilepsy in a family with a new SLC20A2 mutation.
Idiopathic basal ganglia calcification (IBGC) or primary familial brain calcification is a rare genetic condition characterized by an autosomal dominant inheritance pattern and the presence of bilateral calcifications in the basal ganglia, thalami, cerebellum and cerebral subcortical white matter. The syndrome is genetically and phenotypically heterogeneous. Causal mutations have been identified in four genes: SLC20A2, PDGFRB, PDGFB and XPR1. A variety of progressive neurological and psychiatric symptoms have been described, including cognitive impairment, movement disorders, bipolar disorder, chronic headaches and migraine, and epilepsy. Here we describe a family with a novel SLC20A2 mutation mainly presenting with neurological symptoms including cortical myoclonus and epilepsy. While epilepsy, although rare, has been reported in patients with IBGC associated with SLC20A2 mutations, cortical myoclonus seems to be a new manifestation.


Susceptibility of Neisseria meningitidis and Neisseria gonorrhoeae isolates to N-formimidoyl thienamycin.
A diverse collection of 123 meningococcal and 126 gonococcal isolates was tested for susceptibility to N-formimidoyl thienamycin (N-F-thienamycin; MK0787) and to penicillin G (PEN). All of the meningococci were susceptible to both of these, as well as to rifampin. Among gonococci, beta-lactamase-producing strains, which were resistant to PEN, were susceptible to N-F-thienamycin. Among non-beta-lactamase-producing strains, the minimal inhibitory concentrations of N-F-thienamycin and PEN were less than or equal to 1.28 micrograms/ml. Of these, the strains with PEN minimal inhibitory concentrations toward the higher end of the range were likely to be more susceptible to N-F-thienamycin, whereas the strains that were highly susceptible to PEN were likely to have higher minimal inhibitory concentrations of N-F-thienamycin.


Vitamin and mineral supplement use by children and adolescents in the 1999-2004 National Health and Nutrition Examination Survey: relationship with nutrition, food security, physical activity, and health care access.
To determine if vitamin and mineral supplement use among children and adolescents in the United States is associated with nutrition, food security, physical activity, and health care access. Secondary analysis of nationally representative data from the 1999-2004 National Health and Nutrition Examination Survey. Questionnaires, household interviews, and medical examinations. Children and adolescents 2 to 17 years of age (N = 10 828). Vitamin and mineral supplement use in the past month. Demographics, nutrition, food security, physical activity, and health care access. Approximately 34% (SE 1.2) used vitamin and mineral supplements in the past month, with underweight subjects reporting greater intake. Younger age, non-Hispanic white race/ethnicity, being born in the United States, higher milk intake, lower total fat and cholesterol intake, higher dietary fiber intake, higher income, greater food security, lower media/computer use, greater physical activity, lower body mass index, health insurance coverage, better health care access, and better self-reported health were associated with greater use of vitamin and mineral supplements. Such supplements contributed significantly to total daily dietary intakes of vitamins and minerals. A large number of US children and adolescents use vitamin and mineral supplements, which for most may not be medically indicated. Such supplements contribute significantly to total dietary intakes of vitamins and minerals, and studies of nutrition should include their assessment. Since vitamin and mineral supplement users report greater health care access, health care providers may be in a position to provide screening and counseling regarding dietary adequacy and indications for supplement use.


The humoral immune system has a key prognostic impact in node-negative breast cancer.
Estrogen receptor (ER) expression and proliferative activity are established prognostic factors in breast cancer. In a search for additional prognostic motifs, we analyzed the gene expression patterns of 200 tumors of patients who were not treated by systemic therapy after surgery using a discovery approach. After performing hierarchical cluster analysis, we identified coregulated genes related to the biological process of proliferation, steroid hormone receptor expression, as well as B-cell and T-cell infiltration. We calculated metagenes as a surrogate for all genes contained within a particular cluster and visualized the relative expression in relation to time to metastasis with principal component analysis. Distinct patterns led to the hypothesis of a prognostic role of the immune system in tumors with high expression of proliferation-associated genes. In multivariate Cox regression analysis, the proliferation metagene showed a significant association with metastasis-free survival of the whole discovery cohort [hazard ratio (HR), 2.20; 95% confidence interval (95% CI), 1.40-3.46]. The B-cell metagene showed additional independent prognostic information in carcinomas with high proliferative activity (HR, 0.66; 95% CI, 0.46-0.97). A prognostic influence of the B-cell metagene was independently confirmed by multivariate analysis in a first validation cohort enriched for high-grade tumors (n = 286; HR, 0.78; 95% CI, 0.62-0.98) and a second validation cohort enriched for younger patients (n = 302; HR, 0.83; 95% CI, 0.7-0.97). Thus, we could show in three cohorts of untreated, node-negative breast cancer patients that the humoral immune system plays a pivotal role in metastasis-free survival of carcinomas of the breast.


Characteristics of persons convicted for offences relating to animal hoarding in New South Wales.
To highlight the characteristics of persons convicted for offences related to animal hoarding in New South Wales, Australia, document the outcomes of cases and compare them with overseas studies. Retrospective case series. Records of finalised prosecutions for offences relating to animal hoarding between 2005 and 2011 were examined. Data recorded included: the age of each subject at the first offence, sex, postcode, occupation, living conditions, number of charges, number of prosecutions, title of each charge, number and species of live animals, whether animals needed veterinary attention, the medical conditions that the animals suffered, whether dead animals were on the property, how animals were obtained, veterinary and legal costs accrued and case outcomes. The data were analysed to obtain frequencies and relative frequencies for categorical variables and summary statistics for quantitative variables. Observed frequencies were compared using Chi-square test with the expected frequencies calculated based on the Australian Bureau of Statistics data for NSW. The number of persons included was 29. Most were female (72.4%) and 23 were 40-64 years of age at their first offence. Almost one-third identified themselves as breeders, eight as pensioners and four as unemployed. Most resided in inner regional Australia (45%), 28% lived in major cities and 28% lived in outer regional Australia. Dogs were the species hoarded in 80% of cases. Animals requiring veterinary attention were identified in all cases. Dead animals were found on premises in 41.4% of cases. Persons prosecuted for charges relating to animal hoarding in NSW have similar characteristics to those of previous studies, although the outcomes may be different. More farm animals and horses were hoarded in NSW and hoarders in NSW were more likely to live in inner regional and outer regional areas (rural areas) than animal hoarders in the USA.


[The effect of molsidomine on the conjunctival microcirculation in patients with ischemic heart disease].
Sublingual administration of molsidomin in average dose of 3 mg with an interval of 15 and 60 minutes was used in the treatment of 24 patients with ischemic heart disease with stable exertion stenocardia of the II-III functional classes. Course treatment in optimal chosen doses resulted in an improvement of the indices of conjunctival microcirculation and rheological properties of erythrocytes.


Safranal Inhibits HeLa Cell Viability by Perturbing the Reassembly Potential of Microtubules.
Saffron, a spice from Crocus sativus, has been known for its health benefits and medicinal properties. Safranal is a component of saffron and is known for its antioxidant and anticancer properties. In this study, we elucidated a possible tubulin-targeted antiproliferative mechanism of action of safranal. In vitro, the compound perturbed secondary structure of tubulin without altering net microtubule polymer mass. It inhibited HeLa cell viability in a concentration-dependent manner, with minimal damage to cellular microtubules. However, it strongly inhibited recovery of microtubule network after cold-induced disassembly, indicating its ability to interfere with the nucleation potential of tubulin. Further, as the acetylation pattern of the safranal-treated microtubules revealed, unlike many tubulin-targeted agents, the compound did not appear to induce persistent stabilization of microtubules. Our data shows an unusual, tubulin-targeted antiproliferative mechanism of safranal. Copyright © 2017 John Wiley & Sons, Ltd.


Further investigations on the mode of entry of vaccinia virus into cells.
Earlier results indicating that vaccinia virus entered L cells by a process of direct fusion between the virus envelope and the plasma membrane of the cell have been confirmed and extended using immuno-ferritin conjugates to locate virus antigens on the host cell surface. After fusion, components of the virus envelope become rapidly dispersed in the plasma membrane. Fusion has also been observed as the predominant mode of entry of vaccinia virus into HeLa cells.


A double-blind controlled study of gabapentin and baclofen as treatment for acquired nystagmus.
We conducted a double-blind crossover trial comparing gabapentin (up to 900 mg/day) to baclofen (up to 30 mg/day) as therapy for acquired nystagmus in 21 patients. We measured visual acuity and the nystagmus before, and at the end of, 2 weeks on each medication. For a group of 15 patients with acquired pendular nystagmus (APN), visual acuity improved significantly with gabapentin, but not with baclofen. Gabapentin significantly reduced APN median eye speed in all three planes, but baclofen did so only in the vertical plane. In 10 patients with APN, the reduction of nystagmus with gabapentin was substantial and 8 of these elected to continue taking the drug. In 6 patients with downbeat or torsional downbeat nystagmus, changes in median slow-phase eye speed were less consistent with both drugs, either increasing or decreasing, and being dependent on viewing conditions. Only 1 patient showed consistent reduction of median eye speed, and this was achieved by either drug. Our findings suggest that gabapentin may be an effective treatment for many patients with APN and that occasional patients with downbeat nystagmus will respond to gabapentin or baclofen.


Oxidative DNA damage and its repair in rat spleen following subchronic exposure to aniline.
The mechanisms by which aniline exposure elicits splenotoxic response, especially the tumorigenic response, are not well-understood. Splenotoxicity of aniline is associated with iron overload and generation of reactive oxygen species (ROS) which can cause oxidative damage to DNA, proteins and lipids (oxidative stress). 8-Hydroxy-2'-deoxyguanosine (8-OHdG) is one of the most abundant oxidative DNA lesions resulting from ROS, and 8-oxoguanine glycosylase 1 (OGG1), a specific DNA glycosylase/lyase enzyme, plays a key role in the removal of 8-OHdG adducts. This study focused on examining DNA damage (8-OHdG) and repair (OGG1) in the spleen in an experimental condition preceding a tumorigenic response. To achieve that, male Sprague-Dawley rats were subchronically exposed to aniline (0.5 mmol/kg/day via drinking water for 30 days), while controls received drinking water only. Aniline treatment led to a significant increase in splenic oxidative DNA damage, manifested as a 2.8-fold increase in 8-OHdG levels. DNA repair activity, measured as OGG1 base excision repair (BER) activity, increased by approximately 1.3 fold in the nuclear protein extracts (NE) and approximately 1.2 fold in the mitochondrial protein extracts (ME) of spleens from aniline-treated rats as compared to the controls. Real-time PCR analysis for OGG1 mRNA expression in the spleen revealed a 2-fold increase in expression in aniline-treated rats than the controls. Likewise, OGG1 protein expression in the NEs of spleens from aniline-treated rats was approximately 1.5 fold higher, whereas in the MEs it was approximately 1.3 fold higher than the controls. Aniline treatment also led to stronger immunostaining for both 8-OHdG and OGG1 in the spleens, confined to the red pulp areas. It is thus evident from our studies that aniline-induced oxidative stress is associated with increased oxidative DNA damage. The BER pathway was also activated, but not enough to prevent the accumulation of oxidative DNA damage (8-OHdG). Accumulation of mutagenic oxidative DNA lesions in the spleen following exposure to aniline could play a critical role in the tumorigenic process.


Magnetic resonance-guided focused ultrasound surgery treatment of non-spinal intra-articular osteoblastoma: feasibility, safety, and outcomes in a single-center retrospective analysis.
Background: Interventional radiology, thanks to its low invasiveness and possibility to reduce the average time for the patients to come back to their normal activity, is becoming more and more promising and diffused in multiple fields. Employed without needles, MRgFUS is probably the less invasive techniques among the ones belonging to the field of interventional radiology. Purpose: To evaluate safety and effectiveness of MRgFUS in the treatment of a rare and benign, though disabling, bone lesion: intra-articular osteoblastoma. Materials and methods: A retrospective study was carried out on 6 patients (mean, 21 years) treated in the last 2 years with MRgFUS for symptomatic, histologically proved intra-articular osteoblastoma. The main inclusion criterion was the presence of a good acoustic window. The procedures consisted in MR-guided ablation, using high intensity ultrasound beams focused on the target lesion. Spinal anesthesia or peripheral nerve block was used. Clinical (based on pain and functional scales) and imaging follow-up studies were performed up to 1 year after treatment. Complications were recorded. Multiple linear regression and analysis of variance were used to assess correlations. Results: All the procedures were technically successful; no complications were observed. Painful symptomatology decreased of 88% at 6 months and 98% at 12 months (p < 0.0001), and was associated to functional improvement (p = 0.002). MRI and CT controls showed disappearance of all signs of disease and bone inflammation with a marked tendency to bone healing. Conclusion: This study shows the safety and effectiveness of MRgFUS in the treatment of intra-articular osteoblastoma with a good acoustic window.


Clinical Decision-Making in Hypersensitivity Pneumonitis: Diagnosis and Management.
This review provides an updated approach to the diagnosis and management of hypersensitivity pneumonitis (HP). The importance of using a multidisciplinary discussion to increase diagnostic and treatment confidence is emphasized. The role of Bayesian reasoning is highlighted throughout, underscoring the importance of hypothesis generation (differential diagnosis) and diagnostic test interpretation based on the probability of HP. Probability estimates of diagnostic certainty (i.e., a confident versus a working diagnosis) and treatment thresholds are carefully examined.Therapeutically, beyond antigen avoidance and newly available antifibrotic therapy for patients with a progressive fibrosing phenotype; the role, timing, and expected response to anti-inflammatory therapy in individual patients are unanswered questions. Since the evidence and validation of testing generally performed during the diagnostic work-up and longitudinal monitoring of HP is feeble at best, the viewpoints discussed are not intended to resolve current controversies but rather to provide a conceptual framework for evaluating discordant information when evaluating and caring for patients with HP.


Gastro-intestinal and oral microbiome signatures associated with healthy aging.
The human oral and gut microbiomes influence health via competition for a distinct niche in the body with pathogens, via metabolic capabilities that increase host digestive capacity and generate compounds engaged in signaling pathways and modulation of immune system functions. Old age alters our metabolic and regenerative capacity. Following recruitment of 65 human subjects in the age range of 70 to 82, we discerned healthy aging (HA) and non-healthy aging (NHA) cohorts discordant in the occurrence of one or more major diseases: (1) cancer, (2) acute or chronic cardiovascular diseases, (3) acute or chronic pulmonary diseases, (4) diabetes, and (5) stroke or neurodegenerative disorders. We analyzed these cohorts' oral microbiomes (saliva) and gut microbiomes (stool) to assess diversity and identify microbial biomarkers for HA. In contrast to the gut microbiome where no change was observed, we found that the saliva microbiome had higher α-diversity in the HA compared with the NHA group. We observed the genus Akkermansia to be significantly more abundant in the gut microbiota of the HA group. Akkermansia muciniphila is a colonic mucin-degrading bacterium believed to have beneficial effects on gastrointestinal health, particularly in the context of diabetes and obesity. Erysipelotrichaceae UCG-003 was a taxon increased in abundance in the HA cohort. Streptococcus was the only genus observed to be significantly decreased in abundance in both the gut and oral microbiomes of the HA cohort compared with the NHA cohort. Our data support the notion that these microbes are potential probiotics to decrease the risks of non-healthy aging.


CYP1B1, VEGFA, BCL2, and CDKN1A Affect the Development of Chronic Obstructive Pulmonary Disease.
Chronic obstructive pulmonary disease (COPD) is a progressive lung disease characterized by poor airflow. The purpose of this study was to explore the mechanisms involved in the development of COPD. The mRNA expression profile GSE100281, consisting of 79 COPD and 16 healthy samples, was acquired from the Gene Expression Omnibus database. The differentially expressed genes (DEGs) between COPD samples and healthy samples were analyzed using the limma package. Functional enrichment analysis for the DEGs was carried out using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) tool. Furthermore, DEG-compound pairs were predicted using the Comparative Toxicogenomics Database. The KEGG metabolite IDs corresponding to the compounds were also obtained through the MetaboAnalyst pipeline. Based on the diffusion algorithm, the metabolite network was constructed. Finally, the expression levels of key genes were determined using quantitative PCR (qPCR). There were 594 DEGs identified between the COPD and healthy samples, including 242 upregulated and 352 downregulated genes. A total of 696 DEG-compound pairs, such as BCL2-C00469 (ethanol) and BCL2-C00389 (quercetin) pairs, were predicted. CYP1B1, VEGFA, BCL2, and CDKN1A were included in the top 10 DEG-compound pairs. Additionally, 57 metabolites were obtained. In particular, hsa04750 (inflammatory mediator regulation of TRP channels)-C00469 (ethanol) and hsa04152 (AMPK signaling pathway)-C00389 (quercetin) pairs were found in the metabolite network. The results of qPCR showed that the expression of CYP1B1, VEGFA, BCL2, and CDKN1A was consistent with that predicted using bioinformatic analysis. CYP1B1, VEGFA, BCL2, and CDKN1A may play important functions in the development and progression of COPD.


Molecular basis for the resistance of an insect chymotrypsin to a potato type II proteinase inhibitor.
Plants produce a variety of proteinase inhibitors (PIs) that have a major function in defense against insect herbivores. In turn, insects have developed strategies to minimize the effect of dietary PIs on digestion. We have discovered that Helicoverpa larvae that survive consumption of a multidomain serine PI from Nicotiana alata (NaPI) contain high levels of a chymotrypsin that is not inhibited by NaPI. Here we describe the isolation of this NaPI-resistant chymotrypsin and an NaPI-susceptible chymotrypsin from Helicoverpa larvae, together with their corresponding cDNAs. We investigated the mechanism of resistance by mutating selected positions of the NaPI-susceptible chymotrypsin using the corresponding amino acids of the NaPI-resistant chymotrypsin. Four critical residues that conferred resistance to NaPI were identified. Molecular modeling revealed that a Phe-->Leu substitution at position 37 in the chymotrypsin results in the loss of important binding contacts with NaPI. Identification of the molecular mechanisms that contribute to PI resistance in insect digestive proteases will enable us to develop better inhibitors for the control of lepidopteran species that are major agricultural pests worldwide.


Altered gene expression profiling in endometrium: evidence for progesterone resistance.
Progesterone plays an important role in regulating multiple events in the uterus. It controls endometrial proliferation and differentiation, which are important for uterine function. Dysregulation of progesterone signaling leads to impaired physiological functions. Indeed, aberrant expression of progesterone-regulated genes in the endometrium has been implicated in several gynecologic disorders, including endometriosis, polycystic ovarian syndrome (PCOS), and endometrial hyperplasia. Although several investigators have analyzed eutopic endometrial expression of progesterone-target genes, the genesis and consequences of progesterone resistance remain unclear. We review evidence for progesterone resistance in endometrium of women with endometriosis, PCOS, and endometrial hyperplasia, and we identify possible mechanisms associated with reduced progesterone activity in endometrium of (some) women with these gynecologic disorders that have a significant impact on women's health and well-being.


Time to first treatment: The significance of early treatment of exudative age-related macular degeneration.
To determine whether the time span between initial symptoms and treatment with ranibizumab in patients with neovascular age-related macular degeneration has an effect on visual outcome. In this retrospective study, 45 patients with exudative age-related macular degeneration were split into 3 groups depending on the duration of visual symptoms--Group I: <1 month, Group II: 1 month to 6 months, and Group III: >6 months. Best-corrected visual acuity, clinical ophthalmologic examination, and central retinal thickness as measured by optical coherence tomography were recorded at baseline and 2 months later. Fluorescein angiography was performed at baseline. Treatment consisted of 2 intravitreal injections of 1.25 mg of ranibizumab at baseline and after 4 weeks. The mean time span between initial symptoms and treatment was 59 ± 62 days. In all groups, a reduction of retinal thickness was observed. Shorter disease duration, as estimated by persistence of visual symptoms, was correlated with a better visual outcome after treatment. Patients in Group I demonstrated a significant increase in best-corrected visual acuity (P = 0.007). Patients of Group II (P = 0.095) and Group III (P = 0.271) still achieved a visual improvement in best-corrected visual acuity, albeit not significant. The mean change in best-corrected visual acuity was 0.08 ± 0.1 in all patients and was not statistically significant between groups (P = 0.87). Duration of visual symptoms <1 month before treatment is associated with a better visual outcome. Treatment of new-onset wet age-related macular degeneration should be initiated as soon as possible.


Association of Ascending Aortic Dilatation and Long-term Endurance Exercise Among Older Masters-Level Athletes.
Aortic dilatation is frequently encountered in clinical practice among aging endurance athletes, but the distribution of aortic sizes in this population is unknown. It is additionally uncertain whether this may represent aortic adaptation to long-term exercise, similar to the well-established process of ventricular remodeling. To assess the prevalence of aortic dilatation among long-term masters-level male and female athletes with about 2 decades of exercise exposure. This cross-sectional study evaluated aortic size in veteran endurance athletes. Masters-level rowers and runners aged 50 to 75 years were enrolled from competitive athletic events across the United States from February to October 2018. Analysis began January 2019. Long-term endurance exercise. The primary outcome was aortic size at the sinuses of Valsalva and the ascending aorta, measured using transthoracic echocardiography in accordance with contemporary guidelines. Aortic dimensions were compared with age, sex, and body size-adjusted predictions from published nomograms, and z scores were calculated where applicable. Among 442 athletes (mean [SD] age, 61 [6] years; 267 men [60%]; 228 rowers [52%]; 214 runners [48%]), clinically relevant aortic dilatation, defined by a diameter at sinuses of Valsalva or ascending aorta of 40 mm or larger, was found in 21% (n = 94) of all participants (83 men [31%] and 11 women [6%]). When compared with published nomograms, the distribution of measured aortic size displayed a rightward shift with a rightward tail (all P < .001). Overall, 105 individuals (24%) had at least 1 z score of 2 or more, indicating an aortic measurement greater than 2 SDs above the population mean. In multivariate models adjusting for age, sex, body size, hypertension, and statin use, both elite competitor status (rowing participation in world championships or Olympics or marathon time under 2 hours and 45 minutes) and sport type (rowing) were independently associated with aortic size. Clinically relevant aortic dilatation is common among aging endurance athletes, raising the possibility of vascular remodeling in response to long-term exercise. Longitudinal follow-up is warranted to establish corollary clinical outcomes in this population.


Comparable effects of alendronate and strontium ranelate on femur in ovariectomized rats.
This study compared the effects of alendronate (ALN) and strontium ranelate (SR) on bone mineral density (BMD), bone histomorphometry, and biomechanics in ovariectomized (OVX) rats. We randomly assigned 48 3-month-old female Sprague-Dawley rats to four groups: sham, OVX, ALN, and SR. Rats in the OVX, ALN, and SR groups received bilateral OVX. Rats in the ALN and SR groups were orally administrated ALN (7 mg/kg/week) and SR (500 mg/kg/day). Rats in the sham and OVX groups were treated with saline. All treatments continued for 12 weeks. Femoral BMD examination, distal femoral bone histomorphometry analysis, and biomechanical tests at the femoral diaphysis and metaphysis were performed to evaluate the effects of treatments in OVX rats. Results showed that both ALN and SR significantly increased femoral BMD (total femur, diaphyseal BMD, and distal metaphyseal BMD), distal femoral bone histomorphometric parameters (BV/TV, Tb.N, and Tb.Th), and femoral biomechanical parameters (maximum load, failure load, stiffness) compared with the OVX group (P < 0.05). No differences were found between ALN and SR in increasing femoral BMD, distal femoral bone histomorphometric parameters (BV/TV, Tb.N, and Tb.Th), and femoral diaphysis biomechanical parameters (maximum load, failure load, stiffness) (P > 0.05). The SR group was inferior to the ALN group in femoral metaphysis biomechanical parameters (P < 0.05). In conclusion, ALN (7 mg/kg/week) and SR (500 mg/kg/day) have similar effects by increasing BMD, distal femoral bone histomorphometric parameters, and femoral metaphysis biomechanical properties. Although ALN has greater effects than SR on distal femoral metaphysis biomechanical properties, in general, ALN and SR have comparable effects on the femur in OVX rats.


Influence of oxygen on NADH recycling and oxidative stress resistance systems in Lactobacillus panis PM1.
Lactobacillus panis strain PM1 is an obligatory heterofermentative and aerotolerant microorganism that also produces 1,3-propanediol from glycerol. This study investigated the metabolic responses of L. panis PM1 to oxidative stress under aerobic conditions. Growth under aerobic culture triggered an early entrance of L. panis PM1 into the stationary phase along with marked changes in end-product profiles. A ten-fold higher concentration of hydrogen peroxide was accumulated during aerobic culture compared to microaerobic culture. This H2O2 level was sufficient for the complete inhibition of L. panis PM1 cell growth, along with a significant reduction in end-products typically found during anaerobic growth. In silico analysis revealed that L. panis possessed two genes for NADH oxidase and NADH peroxidase, but their expression levels were not significantly affected by the presence of oxygen. Specific activities for these two enzymes were observed in crude extracts from L. panis PM1. Enzyme assays demonstrated that the majority of the H2O2 in the culture media was the product of NADH: H2O2 oxidase which was constitutively-active under both aerobic and microaerobic conditions; whereas, NADH peroxidase was positively-activated by the presence of oxygen and had a long induction time in contrast to NADH oxidase. These observations indicated that a coupled NADH oxidase - NADH peroxidase system was the main oxidative stress resistance mechanism in L. panis PM1, and was regulated by oxygen availability. Under aerobic conditions, NADH is mainly reoxidized by the NADH oxidase - peroxidase system rather than through the production of ethanol (or 1,3-propanediol or succinic acid production if glycerol or citric acid is available). This system helped L. panis PM1 directly use oxygen in its energy metabolism by producing extra ATP in contrast to homofermentative lactobacilli.


Middle cerebral artery infarction: relationship of cavernous carotid artery calcification.
This study was designed to determine whether calcification of the cavernous carotid artery (CCA) is associated with cerebrovascular infarcts in the same way that coronary artery calcification scores indicate myocardial infarctions. We sought to correlate the grade of CCA calcification with infarctions in the middle cerebral artery (MCA) distribution. Nonenhanced brain CT scans of 40 patients with MCA-distribution strokes, 34 with non-MCA-distribution strokes, and 94 age-matched control subjects were reviewed. Circumferential calcification and thickness of calcification were graded for the CCAs on head CT scans. Scores were determined for the left and right CCAs. Mann-Whitney tests and Spearman correlation coefficients were used to detect differences between patients and control subjects and between patients with MCA and those with non-MCA strokes. CCA calcification scores did not significantly differ in the groups compared. The manner in which calcification was scored (by using circumference, thickness, or both) did not affect the results. No difference was noted between scores ipsilateral and those contralateral to the stroke. Circumferential degree or thickness of cavernous artery calcification was not correlated with MCA or non-MCA infarctions. CCA calcification scores did not differ between patients with stroke and those without stroke.


Interaction of Saccharomyces boulardii with Salmonella enterica serovar Typhimurium protects mice and modifies T84 cell response to the infection.
Salmonella pathogenesis engages host cells in two-way biochemical interactions: phagocytosis of bacteria by recruitment of cellular small GTP-binding proteins induced by the bacteria, and by triggering a pro-inflammatory response through activation of MAPKs and nuclear translocation of NF-kappaB. Worldwide interest in the use of functional foods containing probiotic bacteria for health promotion and disease prevention has increased significantly. Saccharomyces boulardii is a non-pathogenic yeast used as a probiotic in infectious diarrhea. In this study, we reported that S. boulardii (Sb) protected mice from Salmonella enterica serovar Typhimurium (ST)-induced death and prevented bacterial translocation to the liver. At a molecular level, using T84 human colorectal cancer cells, we demonstrate that incubation with Sb before infection totally abolished Salmonella invasion. This correlates with a decrease of activation of Rac1. Sb preserved T84 barrier function and decreased ST-induced IL-8 synthesis. This anti-inflammatory effect was correlated with an inhibitory effect of Sb on ST-induced activation of the MAPKs ERK1/2, p38 and JNK as well as on activation of NF-kappaB. Electron and confocal microscopy experiments showed an adhesion of bacteria to yeast cells, which could represent one of the mechanisms by which Sb exerts its protective effects. Sb shows modulating effects on permeability, inflammation, and signal transduction pathway in T84 cells infected by ST and an in vivo protective effect against ST infection. The present results also demonstrate that Sb modifies invasive properties of Salmonella.


Divergent isoenzyme profiles of sylvatic and domiciliary Trypanosoma cruzi in the eastern plains, piedmont, and highlands of Colombia.
Fifty-four stocks of Trypanosoma cruzi from vectors, mammalian reservoirs, and infected humans were characterized by enzyme electrophoresis in starch gels using Brazilian zymodeme reference strains (Z1, X-10; Z2, ESM; Z3, CAN-3) as standards. Colombian stocks were collected in three ecologically and epidemiologically distinct settings. Thirteen enzymes were included in the evaluation. Sixteen different phenotypic profiles or "zymodemes" were evident and generated three groups of closely related stocks: a sylvatic Z1-like group, a domiciliary Z1-like group, and a sylvatic Z3-like group. The number of zymodemes observed in foci of sylvatic transmission was greater than in foci of domiciliary transmission. Modified ecologic conditions associated with agriculture and the consequent reduction of biologic diversity may account for the observed pattern of zymodeme distribution and heterogeneity. The phenotypic similarity between the principal sylvatic group of stocks and domiciliary stocks contrasts with the extensive differences observed between the domestic Z2 zymodeme and sylvatic Z1 and Z3 zymodemes in Brazil and Chile.


Human CD271-positive melanoma stem cells associated with metastasis establish tumor heterogeneity and long-term growth.
Human melanoma is composed of distinct cell types reminiscent of neural crest derivatives and contains multipotent cells that express the neural crest stem cell markers CD271(p75(NTR)) and Sox10. When isolated from solid tumors by using a method that leaves intact cell surface epitopes, CD271-positive, but not CD271-negative, cells formed tumors on transplantation into nude or nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. These tumors fully mirrored the heterogeneity of the parental melanoma and could be passaged more than 5 times. In contrast, in more immunocompromised NOD/SCID/IL2rγ(null) mice, or in natural killer cell-depleted nude or NOD/SCID mice, both CD271-positive and CD271-negative tumor cell fractions established tumors. However, tumors resulting from either fraction did not phenocopy the parental tumors, and tumors derived from the CD271-negative cell fraction could not be passaged multiple times. Together, our findings identify CD271-positive cells as melanoma stem cells. Our observation that a relatively high frequency of CD271/Sox10-positive cells correlates with higher metastatic potential and worse prognosis further supports that CD271-positive cells within human melanoma represent genuine cancer stem cells.


Familial and Hereditary Forms of Primary Hyperparathyroidism.
Individuals with a familial predisposition to the development of parathyroid tumors constitute a small minority of all patients with primary hyperparathyroidism (PHPT). These familial syndromes exhibit Mendelian inheritance patterns and the main causative genes in most families have been identified. They include multiple endocrine neoplasia (MEN; types 1, 2A, and 4), hyperparathyroidism-jaw tumor (HPT-JT) syndrome, familial isolated hyperparathyroidism, familial hypocalciuric hypercalcemia (FHH), and neonatal severe PHPT. Each MEN type is associated with the various combinations of specific tumors. MEN1 is characterized by the occurrence of parathyroid, enteropancreatic, and pituitary tumors; MEN2A is characterized by medullary thyroid carcinoma and pheochromocytoma, and MEN4 is characterized by a pathological spectrum similar to that of MEN1 in association with tumors of the adrenal, kidney, and reproductive organs. HPT-JT is characterized by PHPT, ossifying fibromas of maxillary bones, kidney disease, and uterine neoplasias. The prompt diagnosis of these diseases is of great importance for planning appropriate surveillance of the mutant carriers and correct surgical management. The search for mutation is also useful for the identification of the family members who do not carry the mutation and can avoid unnecessary biochemical and instrumental evaluations. Surgery remains the treatment of choice in all familial forms except FHH.