Effects of Alu elements on global nucleosome positioning in the human genome.
Understanding the genome sequence-specific positioning of nucleosomes is essential to understand various cellular processes, such as transcriptional regulation and replication. As a typical example, the 10-bp periodicity of AA/TT and GC dinucleotides has been reported in several species, but it is still unclear whether this feature can be observed in the whole genomes of all eukaryotes. With Fourier analysis, we found that this is not the case: 84-bp and 167-bp periodicities are prevalent in primates. The 167-bp periodicity is intriguing because it is almost equal to the sum of the lengths of a nucleosomal unit and its linker region. After masking Alu elements, these periodicities were greatly diminished. Next, using two independent large-scale sets of nucleosome mapping data, we analyzed the distribution of nucleosomes in the vicinity of Alu elements and showed that (1) there are one or two fixed slot(s) for nucleosome positioning within the Alu element and (2) the positioning of neighboring nucleosomes seems to be in phase, more or less, with the presence of Alu elements. Furthermore, (3) these effects of Alu elements on nucleosome positioning are consistent with inactivation of promoter activity in Alu elements. Our discoveries suggest that the principle governing nucleosome positioning differs greatly across species and that the Alu family is an important factor in primate genomes.


Severity of RSV infection in Southern European elderly patients during two consecutive winter seasons (2017-2018).
In Europe, the respiratory syncytial virus (RSV) surveillance system is very heterogeneous and there is growing evidence of the importance of RSV infections resulting in hospitalization of elderly patients. The aim of this study was to assess the severity of RSV infection in the elderly living in the aged Southern European countries. We conducted a retrospective study of elderly patients ( ≥65-year old) admitted for laboratory-confirmed RSV infection in three tertiary hospitals in Portugal, Italy, and Cyprus over two consecutive winter seasons (2017-2018). Uni-multivariable analyses were carried out to evaluate the effect of clinical variables on radiologically confirmed pneumonia, use of noninvasive ventilation (NIV), and in-hospital death (IHD). A total of 166 elderly patients were included. Pneumonia was evident in 29.5%. NIV was implemented in 16.3%, length of stay was 11.8 ± 12.2 days, and IHD occurred in 12.1%. Multivariable analyses revealed that the risk of pneumonia was higher in patients with chronic kidney disease (CKD) (odds ratio [OR]: 2.57; 95% confidence interval [CI]: 1.12-5.91); the use of NIV was higher in patients with obstructive sleep apnea or obesity hypoventilation syndrome (OSA or OHS) (OR: 5.38; 95% CI: 1.67-17.35) and CKD (OR: 2.52; 95% CI: 1.01-6.23); the risk of IHD was higher in males (OR: 3.30; 95% CI: 1.07-10.10) and in patients with solid neoplasm (OR: 9.06; 95% CI: 2.44-33.54) and OSA or OHS (OR: 8.39; 95% CI: 2.14-32.89). Knowledge of factors associated with RSV infection severity may aid clinicians to set priorities and reduce disease burden. Development of effective antiviral treatment and vaccine against RSV is highly desirable.


2019 novel Coronavirus: current knowledge, cardiovascular implications and management.
From first cases reported on December 31, 2019, in Wuhan, Hubei-China, SARS-CoV2 has spread worldwide and finally the World Health Organization declared the pandemic status. We summarize what makes SARS-CoV2 different from previous highly pathogenic coronaviruses and why it is so contagious, with focus on its clinical presentation and diagnosis, which is mandatory to start the appropriate management and reduce the transmission. As far as infection pathophysiology is still not completely clarified, this review focuses also on the cardiovascular (CV) implication of COVID-19 and the capability of this virus to cause direct myocardial injury, myocarditis and other CV manifestations. Furthermore, we highlight the relationship between the virus, enzyme ACE2 and ACE inhibitors. Clinical management involves the intensive care approach with intubation and mechanical ventilation in the most serious cases and drug therapy with several apparently promising old and new molecules. Aim of this review is then to summarize what is actually known about the SARS-CoV2 and its cardiovascular implications.


A prospective randomized trial of buffy coat versus CD34-selected autologous bone marrow support in high-risk breast cancer patients receiving high-dose chemotherapy.
High-dose chemotherapy with hematopoietic progenitor cell support is administered increasingly to selected categories of patients with high-risk malignancies. Bone marrow and/or peripheral blood progenitor cells (PBPCs) are commonly cryopreserved with the cryoprotectant dimethyl sulfoxide (DMSO), which can cause a variety of systemic side effects when the graft is thawed and infused. The progenitor cells thought to be responsible for hematopoietic recovery express the CD34 antigen and constitute 1% to 3% of the marrow cells and 0.5% of the PBPC fraction. Transplantation of a CD34(+) graft would markedly reduce the volume and thus the amount of DMSO required, thereby decreasing the infusion-related toxicities. In this study, 89 high-risk breast cancer patients received high-dose therapy and were randomized to receive an autologous CD34(+) marrow graft (Arm A) versus a standard buffy coat fraction (Arm B). After marrow infusion, significant increases in diastolic and systolic blood pressure, as well as significant decreases in heart rate, were documented in Arm B compared to Arm A patients (P < .001). None of the patients in Arm A experienced any clinically serious adverse events associated with the marrow infusion compared to 6% of the Arm B patients. The median time to neutrophil engraftment was 13 days for Arm A and 11 days for Arm B patients (P = .218). The median time to platelet engraftment was 27 days for Arm A and 20 days for Arm B patients (0.051). There were no other significant differences between the two arms of the study with respect to thrombocytopenia-related complications or immune function reconstitution. Additionally, patients on Arm A who received >/=1.2 x 10(6) CD34(+) cells/kg had no delay in platelet recovery (22 days), compared to patients on Arm B, who also received greater than 1.2 x 10(6) CD34(+) cells/kg (20 days) (P = .604). In conclusion, this prospective randomized study demonstrates that breast cancer patients who receive high-dose therapy with autologous CD34(+) marrow support have reduced marrow infusion-related toxicity, comparable time to neutrophil engraftment and immune function recovery posttransplant, and for those who receive <1.2 x 10(6) CD34(+) cells/kg, comparable time to platelet engraftment compared to women who receive buffy coat fractions of marrow.


K88-mediated binding of Escherichia coli outer membrane fragments to porcine intestinal epithelial cell brush borders.
We have examined the interactions between various radiolabeled membrane fractions obtained from an enterotoxigenic Escherichia coli strain and brush borders isolated from porcine intestinal epithelial cells. Outer membrane fragments containing the K88 attachment factor bound tightly to brush borders, whereas cytoplasmic membrane vesicles did not. Three different types of outer membrane preparations were tested: (i) cellular outer membranes isolated from lysozyme spheroplasts, (ii) medium vesicles or outer membrane fragments released into the medium during growth, and (iii) periplasmic vesicles, or outer membrane fragments which were released from the cells during spheroplast formation and were therefore isolated in the periplasmic fraction. Of these fractions, which were heterogeneous, it was always the outer membrane subfraction which bound tightly to brush borders. This binding, which was K88 dependent, may have some physiological significance in view of the association between outer membrane fragments and enterotoxin. Thus, released outer membrane fragments equipped with attachment factors may function as enterotoxin carriers which increase the efficiency with which enterotoxin can be delivered to intestinal epithelial cells.


Excessive Sedation as a Risk Factor for Delirium: A Comparison between Two Cohorts of ARDS Critically Ill Patients with and without COVID-19.
Excessive sedation is associated with poor outcome in critically ill acute respiratory distress syndrome (ARDS) patients. Whether this prognostic effect varies among ARDS patients with and without COVID-19 has yet to be determined. We compared the prognostic value of excessive sedation—in terms of delirium, length of stay in intensive care unit (ICU-LOS) and ICU mortality—between COVID-19 and non-COVID-19 critically ill ARDS patients. This was a second analysis of prospectively collected data in four European academic centers pertaining to 101 adult critically ill ARDS patients with and without COVID-19 disease. Depth of sedation (DOS) and delirium were monitored through processed electroencephalogram (EEG) and the Confusion Assessment Method for ICU (CAM-ICU). Our main exposure was excessive sedation and how it relates to the presence of delirium, ICU-LOS and ICU mortality. The criterion for excessive sedation was met in 73 (72.3%) patients; of these, 15 (82.2%) and 58 (69.1%) were in non-COVID-19 and COVID-19 ARDS groups, respectively. The criteria of delirium were met in 44 patients (60.3%). Moreover, excessive sedation was present in 38 (86.4%) patients with delirium (p < 0.001). ICU death was ascertained in 41 out of 101 (41.0%) patients; of these, 37 (90.2%) had excessive sedation (p < 0.001). The distribution of ICU-LOS among excessive-sedated and non-sedated patients was 22 (16−27) vs. 14 (10.5−19.5) days (p < 0.001), respectively. In a multivariable framework, excessive sedation was independently associated with the development of delirium (p = 0.001), increased ICU mortality (p = 0.009) and longer ICU-LOS (p = 0.000), but only in COVID-19 ARDS patients. Independent of age and gender, excessive sedation might represent a risk factor for delirium in COVID-19 ARDS patients. Similarly, excessive sedation shows to be an independent predictor of ICU-LOS and ICU mortality. The use of continuous EEG-based depth of sedation (DOS) monitoring and delirium assessment in critically ill COVID-19 patients is warranted.


Positive association of serum prolactin concentrations with all-cause and cardiovascular mortality.
Increased serum prolactin (PRL) concentrations have been associated with adverse cardiovascular risk profiles, but the relation between PRL and mortality risk is unknown. We evaluated 3929 individuals (1946 men and 1983 women) aged 20-81 (mean 50.3 years) from the population-based Study of Health in Pomerania (SHIP). Associations of continuous [per standard deviation (SD) increase] and categorized (sex-specific tertiles) serum PRL concentrations with all-cause and cause-specific mortality were analysed separately for men and women by age- and multivariable-adjusted Cox regression models. During a median follow-up period of 10.1 years (38 231 person-years), 419 deaths (10.7%), 132 cardiovascular deaths (3.4%), and 152 cancer deaths (3.9%) were observed. After multivariable adjustment, we observed a positive association of PRL with all-cause mortality in men and women [hazard ratio (HR) per SD increase: 1.17, 95% confidence interval (CI): 1.07-1.29 and HR: 1.22, 95% CI: 1.03-1.46, respectively]. Similarly, individuals with PRL concentrations in the highest tertile (when compared with lowest PRL tertile) experienced the highest mortality risk (men: HR, 1.75; 95% CI, 1.32-2.32; women: HR, 1.66; 95% CI, 1.08-2.56), with a significant trend across PRL tertiles (P- for trend <0.05). Cause-specific mortality analyses yielded similar associations for cardiovascular death in both sexes, but for cancer death only in men. This is the first study to report an independent positive association of PRL concentrations with all-cause and cardiovascular mortality. Further studies are required to confirm our findings and to elucidate the potential role of PRL as a useful biomarker of cardiovascular risk and mortality assessment.


Risk of bile duct carcinogenesis after excision of extrahepatic bile ducts in pancreaticobiliary maljunction.
A reflux of pancreatic juice into the biliary tract caused by pancreaticobiliary maljunction (PBM) has been considered important in the development of biliary tract carcinogenesis in choledochal cysts. We excised extrahepatic bile ducts in patients with choledochal cysts to terminate the reflux of pancreatic juice. We investigated whether this surgery could stop the development of the residual bile duct carcinoma. Fifty-six patients with a diagnosis of PBM with choledochal dilatation underwent surgical excision of extrahepatic bile ducts. We applied a person-year method to compare the relative risks (observed number/expected number) of biliary tract carcinoma before and after surgery. In 3 patients, bile duct carcinoma developed in residual dilated segments 19 years 6 months, 8 years 8 months, and 2 years 5 months, respectively, after surgery. Although the relative risk in the post-surgery group was slightly decreased by surgery, it was still high compared with that of the general population. The incidence of bile duct carcinoma is still high, even after excision of extrahepatic bile ducts in PBM patients with choledochal dilatation. For these patients, careful long-term follow-up is necessary, especially after operations that leave the dilated bile ducts, such as cases of Todani's type IV-A.


Phylogenetic analyses of the constituents of Type III protein secretion systems.
Multicomponent Type III protein secretion systems transfer gram-negative bacterial virulence factors directly from the bacterial cytoplasm to the cytoplasm of a host eukaryotic cell in a process that may involve a single energy-coupled step. Extensive evidence supports the conclusion that the genetic apparatuses that encode these systems have been acquired independently by different gram-negative bacteria, presumably by lateral transfer. In this paper we conduct phylogenetic analyses of currently sequenced constituents of these systems and their homologues. The results reveal the relative relatedness of these systems and show that they evolved with little or no exchange of constituents between systems. This fact suggests that horizontal transmission of the genes encoding these systems always occurred as a unit without the formation of hybrid gene clusters. Moreover, homologous flagellar proteins show phylogenetic clustering that suggests that the flagellar systems and Type III protein secretory systems diverged from each other following very early duplication of a gene cluster sharing many (but not all) genes. Phylogenies of most or all of the flagellar proteins follow those of the source organisms with little or no lateral gene transfer suggesting that homologous flagellar proteins are true orthologues. We suggest that the flagellar apparatus was the evolutionary precursor of Type III protein secretion systems.


Multivariate statistical study of heavy metal enrichment in sediments of the Pearl River Estuary.
The concentrations and chemical partitioning of heavy metals in the sediment cores of the Pearl River Estuary were studied. Based on Pearson correlation coefficients and principal component analysis results, Al was selected as the concentration normalizer for Pb, while Fe was used as the normalizing element for Co, Cu, Ni and Zn. In each profile, sections with metal concentrations exceeding the upper 95% prediction interval of the linear regression model were regarded as metal enrichment layers. The heavy metal accumulation mainly occurred at sites in the western shallow water areas and east channel, which reflected the hydraulic conditions and influence from riparian anthropogenic activities. Heavy metals in the enrichment sections were evaluated by a sequential extraction method for possible chemical forms in sediments. Since the residual, Fe/Mn oxides and organic/sulfide fractions were dominant geochemical phases in the enriched sections, the bioavailability of heavy metals in sediments was generally low. The 206Pb/207Pb ratios in the metal-enriched sediment sections also revealed the influence of anthropogenic sources. The spatial distribution of cumulative heavy metals in the sediments suggested that the Zn and Cu mainly originated from point sources, while the Pb probably came from non-point sources in the estuary.


Tailored Systemic Therapy for Colorectal Cancer Liver Metastases.
Liver metastases are the most common site of metastatic spread in colorectal cancer. Current treatment approaches involve effective systemic therapies in combination with surgical and/or interventional strategies. Multimodal strategies greatly improved clinical outcomes of patients with metastatic colorectal cancer over the last decades. Identification of predictive and prognostic biomarkers helped to comprehensively refine individual targeted treatment approaches and resulted in median overall survival rates of 30 months or longer. Current guidelines, thus, recommend treatment selection according to patients' performance status, tumor localization and stage as well as the tumor's molecular and genetic status. Here, we outline the latest developments in molecular decision-making for patients with upfront resectable, potentially or initially unresectable and non/never-resectable colorectal cancer liver metastases.


Negative-ion chemical ionization gas chromatography-mass spectrometry assay for enantioselective measurement of amphetamines in oral fluid: application to a controlled study with MDMA and driving under the influence cases.
Enantioselective analysis of amphetamine (AM), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyethylamphetamine (MDEA) helps interpret toxicological results. Methods have been described for various matrices, but so far not for oral fluid, a matrix of increasing importance in testing for drugs of abuse, especially in the context of driving under the influence of drugs (DUID). After dilution with 200 microL carbonate buffer (pH 9), oral fluid samples (10-50 microL) were derivatized with S-heptafluorobutyrylprolyl chloride. The resulting diastereomers were extracted into 100 microL of cyclohexane, separated by gas chromatography (HP-5MS column), and detected by mass spectrometry in the negative-ion chemical ionization mode (GC-NICI-MS). The method was validated and applied to samples from a controlled study with MDMA and from authentic DUID cases. The derivatized AM, MA, MDA, MDMA, and MDEA enantiomers were well separated from each other. The method was linear from 5-250 microg/L per enantiomer of MDA and from 25-1250 microg/L per enantiomer of AM, MA, MDMA, and MDEA. With the exception of MDEA, analytical recoveries, repeatability, and intermediate precision were within required limits. The analyte concentrations and enantiomer ratios in the application samples correlated only weakly with corresponding published plasma data. This sensitive, reliable, and fast GC-NICI-MS assay enantioselectively measures AM, MA, MDA, and MDMA in oral fluid samples. Prediction of plasma concentrations and enantiomer ratios from respective oral fluid data is not possible.


Autolysis of Clostridium acetobutylicum ATCC 824.
The optimum conditions for autolysis of Clostridium acetobutylicum ATCC 824 were determined. Autolysis was optimal at pH 6.3 and 55 degrees C in 0.1 M-sodium acetate/phosphate buffer. The ability of cells to autolyse decreased sharply at the end of the exponential phase of growth. Lysis was stimulated by monovalent cations and compounds that complex divalent cations, and inhibited by divalent cations. The autolysin of C. acetobutylicum, which was mainly cytoplasmic, was purified to homogeneity and characterized as a muramidase. The enzyme was identical to the extracellular muramidase in terms of M(r), isoelectric point and NH2-terminal amino acid sequence. The autolysin was inhibited by lipoteichoic acids and cardiolipin but not by phosphatidylethanolamine and phosphatidylglycerol. A mechanism of regulation and fixation involving lipoteichoic acid, cardiolipin and divalent cations is proposed.


Diet and lifestyle factors associated with premenstrual symptoms in a racially diverse community sample: Study of Women's Health Across the Nation (SWAN).
We sought to determine if the frequency of reported physical or emotional premenstrual symptoms (PMSx) was associated with (1) dietary intake of phytoestrogens, fiber, fat, or calcium, (2) consumption of alcohol or caffeine, (3) active or passive smoke exposure or lack of physical exercise, and (4) race/ethnicity or socioeconomic status. A cross-sectional analysis was conducted of PMSx and demographic and lifestyle factors reported at baseline in the multiethnic sample of 3302 midlife women in the Study of Women's Health Across the Nation (SWAN). Stepwise multiple logistic regression analyses were performed for the overall sample and for each racial/ethnic group for each of five PMSx groupings. Most dietary factors were not related to PMSx. Fat intake was negatively associated with craving and bloating (adjusted odds ratio [AOR] = 0.56, p = 0.024), and fiber intake was positively associated with breast pain (AOR = 1.39, p = 0.037). Alcohol intake was negatively associated with anxiety and mood changes (AOR = 0.63, p = 0.045) and headaches (AOR = 0.50, p = 0.009). Current smoking (AOR = 1.60, p = 0.028) and passive smoke exposure (AOR = 1.56, p = 0.050) were positively associated with cramps and back pain. Symptom reporting differed significantly by race/ethnicity. PMSx were also associated with comorbidities, early perimenopausal status, depressive symptoms, and symptom sensitivity. We found little evidence to support a role for diet in PMSx reporting. However, alcohol intake was positively associated with premenstrual anxiety and mood changes, and active and passive smoke exposure was associated with a number of PMSx. Ethnic differences in symptom reporting and associations of comorbidities, early perimenopausal status, depressive symptoms, and symptom sensitivity with reported PMSx were also observed.


Combat urologic trauma in US military overseas contingency operations.
This article reports on the occurrences and patterns of genitourinary (GU) trauma in the contemporary high-intensity conflict of the overseas contingency operations (OCOs). The Joint Theater Trauma Registry was queried for all US military members who received treatment for GU wounds and concomitant injuries sustained in OCOs for >75 months between October 2001 and January 2008. Of the 16,323 trauma admissions annotated in the Joint Theater Trauma Registry, 819 (5%) had one or more GU injuries. Of the GU casualties, 90% were sustained in Iraq and 65% were because of explosions. The average casualty age was 26 years (range, 18-58 years) and 98.5% were men. There were 887 unique GU injuries distributed as follows: scrotum, 257 (29.0%); kidney, 203 (22.9%); bladder, 189 (21.3%); penis, 126 (14.2%); testicle, 81 (9.1%); ureter, 24 (2.7%); and urethra, 7 (0.8%). Of the 203 patients with kidney injuries, 22% went to the operating room with 31 patients having nephrectomies. There were 189 bladder injuries with 26 patients (14%) having concomitant pelvic fractures. This is the largest report of GU injuries during any military conflict. The distribution and percentage of casualties with GU injuries in the OCO are similar to those of previous conflicts. Consideration should be given to personnel protective equipment for the areas associated with GU injuries and predeployment training directed at the care of these injuries.


Protein oxidation and age-dependent alterations in calcium homeostasis.
Alterations in the capacity to maintain normal calcium homeostasis have been suggested to underlie the reduced cellular function characteristic of the aging process, and to predispose the senescent organism to a host of diverse pathologies including cancer, heart disease, and a range of muscle and neurodegenerative diseases. Therefore, critical to the eventual treatment of many age-related diseases has been the identification of both post-translational modifications and the underlying structural changes that result in an age-related decline in the function of critical calcium regulatory proteins. In brain, multiple methionines within the calcium signaling protein calmodulin (CaM) are oxidized to their corresponding methionine sulfoxides during aging, resulting in an inability to activate a range of target proteins, including the plasma membrane (PM) Ca-ATPase involved in the maintenance of the low intracellular calcium levels necessary for intracellular signaling. Likewise, changes in the transport activity of the PM-Ca-ATPase occur during aging. In muscle, the function of the SERCA2a isoform of the Ca-ATPase within the sarcoplasmic reticulum (SR) declines during aging as a result of the nitration of selected tyrosines. The age-related loss-of-function of these critical calcium regulatory proteins are consistent with observed increases in intracellular calcium levels within senescent cells. A possible regulatory role for these post-translational modifications is discussed, since they have the potential to be reversed following the restoration of normal cellular redox conditions by intracellular repair enzymes that are specific for these post-translational modifications. It is suggested that the reversible oxidation of critical calcium regulatory proteins within excitable cells by reactive oxygen species functions to enhance cellular survival under conditions of oxidative stress by reducing the energy expenditure within excitable cells. Thus, a diminished ability to efficiently generate cellular ATP may ultimately underlie the loss of calcium homeostasis and cellular function during aging.


Trigeminal neuralgia caused by a persistent primitive trigeminal artery variant passing through Meckel's cavity: a case report.
Persistent primitive trigeminal artery variant (PPTAv) is a rare remnant of the primitive intracranial embryonic anastomotic arteries, and its persistence has an unknown etiology. Trigeminal neuralgia attributed to a PPTAv passing through Meckel's cavity is extremely uncommon. A 73-year-old woman presented with right-sided facial pain for 10 years that had failed to respond to medication. Magnetic resonance angiography suggested the presence of a PPTAv compressing the trigeminal nerve, as the abnormal artery originated from the right internal carotid artery. During microvascular decompression (MVD), the offending vessel was inferred to be a PPTAv, as it continued to become the anterior inferior cerebellar artery after passing through Meckel's cavity. Postoperative computed tomography angiography showed the PPTAv continuing posteriorly as the anterior inferior cerebellar artery and supplying the cerebellar hemisphere, which confirmed the intraoperative judgment. The pain resolved after MVD and has not recurred in 12 months of follow-up. MVD is the best surgical choice for trigeminal neuralgia combined with a PPTAv. For patients with neurovascular conflicts, particularly those with suspected vascular variations, preoperative imaging examinations play a critical role in meticulously evaluating the anatomical locations of the nerves and blood vessels. Semilunar puncture (for radiofrequency ablation or percutaneous balloon compression) is contraindicated in patients with a PPTAv.


Cross-reactive human B cell and T cell epitopes between influenza A and B viruses.
Influenza A and B viruses form different genera, which were originally distinguished by antigenic differences in their nucleoproteins and matrix 1 proteins. Cross-protection between these two genera has not been observed in animal experiments, which is consistent with the low homology in viral proteins common to both viruses except for one of three polymerase proteins, polymerase basic 1 (PB1). Recently, however, antibody and CD4+ T cell epitopes conserved between the two genera were identified in humans. A protective antibody epitope was located in the stalk region of the surface glycoprotein, hemagglutinin, and a CD4+ T cell epitope was located in the fusion peptide of the hemagglutinin. The fusion peptide was also found to contain antibody epitopes in humans and animals. A short stretch of well-conserved peptide was also identified in the other surface glycoprotein, neuraminidase, and antibodies binding to this peptide were generated by peptide immunization in rabbits. Although PB1, the only protein which has relatively high overall sequence homology between influenza A and B viruses, is not considered an immunodominant protein in the T cell responses to influenza A virus infection, amino acid sequence comparisons show that a considerable number of previously identified T cell epitopes in the PB1 of influenza A viruses are conserved in the PB1 of influenza B viruses. These data indicate that B and T cell cross-reactivity exists between influenza A and B viruses, which may have modulatory effects on the disease process and recovery. Although the antibody titers and the specific T cell frequencies induced by natural infection or standard vaccination may not be high enough to provide cross protection in humans, it might be possible to develop immunization strategies to induce these cross-reactive responses more efficiently.


C4-dicarboxylates sensing mechanism revealed by the crystal structures of DctB sensor domain.
C(4)-dicarboxylates are the major carbon and energy sources during the symbiotic growth of rhizobia. Responses to C(4)-dicarboxylates depend on typical two-component systems (TCS) consisting of a transmembrane sensor histidine kinase and a cytoplasmic response regulator. The DctB-DctD system is the first identified TCS for C(4)-dicarboxylates sensing. Direct ligand binding to the sensor domain of DctB is believed to be the first step of the sensing events. In this report, the water-soluble periplasmic sensor domain of Sinorhizobium meliloti DctB (DctBp) was studied, and three crystal structures were solved: the apo protein, a complex with C(4) succinate, and a complex with C(3) malonate. Different from the two structurally known CitA family of carboxylate sensor proteins CitA and DcuS, the structure of DctBp consists of two tandem Per-Arnt-Sim (PAS) domains and one N-terminal helical region. Only the membrane-distal PAS domain was found to bind the ligands, whereas the proximal PAS domain was empty. Comparison of DctB, CitA, and DcuS suggests a detailed stereochemistry of C(4)-dicarboxylates ligand perception. The structures of the different ligand binding states of DctBp also revealed a series of conformational changes initiated upon ligand binding and propagated to the N-terminal domain responsible for dimerization, providing insights into understanding the detailed mechanism of the signal transduction of TCS histidine kinases.


Genetic variability of the murine creatine kinase B gene locus and related pseudogenes in different inbred strains of mice.
The role of genetic variation in isoenzyme gene families is often poorly appreciated. We report here on the determination of DNA sequences and typing of genetic variability in four creatine kinase B (CKB) gene loci in different inbred strains of mice. The unique functional murine CKB gene was found to be nearly identical to the previously characterised rat and human sequences in both size and exon-intron structure. In this gene, approximately 0.5% allelic nucleotide positions as well as the lengths of simple A-rich and [TG]n repetitive elements located at the 5' and 3' sides of the transcribed segment, differed between inbred strains of mice. Preliminary experiments suggest that this sequence divergence is of importance for design of gene targeting strategies involving homologous DNA recombination. The three additional CKB-like gene loci in mice all had the characteristics of processed pseudogenes. By Southern blot analysis we could demonstrate that both the type and number of pseudogenes differed between inbred strains. Analysis of the CKB gene sequences enabled us to speculate about the evolutionary history of this highly polymorphic subfamily of genes.


Perspective beyond cancer genomics: bioenergetics of cancer stem cells.
Although the notion that cancer is a disease caused by genetic and epigenetic alterations is now widely accepted, perhaps more emphasis has been given to the fact that cancer is a genetic disease. It should be noted that in the post-genome sequencing project period of the 21st century, the underlined phenomenon nevertheless could not be discarded towards the complete control of cancer disaster as the whole strategy, and in depth investigation of the factors associated with tumorigenesis is required for achieving it. Otto Warburg has won a Nobel Prize in 1931 for the discovery of tumor bioenergetics, which is now commonly used as the basis of positron emission tomography (PET), a highly sensitive noninvasive technique used in cancer diagnosis. Furthermore, the importance of the cancer stem cell (CSC) hypothesis in therapy-related resistance and metastasis has been recognized during the past 2 decades. Accumulating evidence suggests that tumor bioenergetics plays a critical role in CSC regulation; this finding has opened up a new era of cancer medicine, which goes beyond cancer genomics.


Direct Thrombin Inhibitors Prevent Left Atrial Remodeling Associated With Heart Failure in Rats.
The present study tested the hypothesis that thrombin participates in formation of left atrial remodeling and that direct oral anticoagulants, such as direct thrombin inhibitors (DTIs), can prevent its progression. In a rat model of heart failure associated with left atrial dilation, we found that chronic treatment with DTIs reduces the atrial remodeling and the duration of atrial fibrillation (AF) episodes induced by burst pacing by inhibiting myocardial hypertrophy and fibrosis. In addition to the prevention of thromboembolism complicating AF, DTIs may be of interest to slow down the progression of the arrhythmogenic substrate.


Chromatographic characterization of adsrenocorticotrophin in human plasma.
A chromatographic procedure has been developed for the characterization of ACTH- and lipotrophin- (LPH) related peptides in human plasma under acid-dissociating conditions to minimize artifacts of protein binding. The recovery and sensitivity of this method permits identification of ACTH at normal physiological levels in the circulation. Plasma profiles obtained from normal subjects and patients with pituitary dependent Cushing's disease, Addison's disease and Nelson's syndsrome showed only one significant peak of ACTH activity eluting in the position of purified native human 1--39 ACTH. However, the plasma profiles obtained from all the patients with the ectopic ACTH syndrome demonstrated a second peak of immunoreactive larger-molecular-weight ACTH and in some plasma samples this was the only form of ACTH observed. This larger-molecular-weight ACTH eluted midway between the void volume and 1--39 ACTH and co-eluted with a protein marker of molecular weight 22 000.


Heterologous Expression of Salvia miltiorrhiza MicroRNA408 Enhances Tolerance to Salt Stress in Nicotiana benthamiana.
MicroRNAs (miRNAs) are a class of endogenous small RNAs that regulate the expression of target genes post-transcriptionally; they are known to play major roles in development and responses to abiotic stress. MicroRNA408 (miR408) is a conserved small RNA in plants; it was reported that miR408 genes were involved in abiotic stress in Arabidopsis. However, miR408 in Salvia miltiorrhiza has been rarely investigated. In this study, we cloned Sm-MIR408, the miR408 precursor sequence, and its promoter sequence from S. miltiorrhiza and the role in tolerance to salt stress is described. The effects of salt stress on miR408 expression were studied by using β-glucuronidase (GUS) staining. Our data indicated that transgenic tobacco overexpressing Sm-MIR408 promoted seed germination and reduced the accumulation of reactive oxygen species under salt stress. Transcript levels of antioxidative genes, i.e., NbSOD, NbPOD, and NbCAT, and their enzyme activities increased in salinity-stressed transgenic tobacco plants, suggesting a better antioxidant system to cope the oxidative damage caused by salinity stress. Taken together, these findings indicated that miR408 functions in positive responses to salt tolerance in tobacco.


Hepatobiliary and pancreatic ascariasis.
Ascariasis is a helminthic infection of global distribution with more than 1.4 billion persons infected throughout the world. The majority of infections occur in the developing countries of Asia and Latin America. Of 4 million people infected in the United States, a large percentage are immigrants from developing countries. Ascaris-related clinical disease is restricted to subjects with heavy worm load, and an estimated 1.2 to 2 million such cases, with 20,000 deaths, occur in endemic areas per year. More often, recurring moderate infections cause stunting of linear growth, cause reduced cognitive function, and contribute to existing malnutrition in children in endemic areas. HPA is a frequent cause of biliary and pancreatic disease in endemic areas. It occurs in adult women and can cause biliary colic, acute cholecystitis, acute cholangitis, acute pancreatitis, and hepatic abscess. RPC causing hepatic duct calculi is possibly an aftermath of recurrent biliary invasion in such areas. Ultrasonography can detect worms in the biliary tract and pancreas and is a useful noninvasive technique for diagnosis and follow-up of such patients. ERCP can help diagnose biliary and pancreatic ascariasis, including ascaris in the duodenum. Also, ERCP can be used to extract worms from the biliary and pancreatic ducts when indicated. Pyrantel pamoate, mebendazole, albendazole and levamisole are effective drugs and can be used for mass therapy to control ascariasis in endemic areas.


Glutathione depletion-induced thymidylate insufficiency for DNA repair synthesis.
Dietary methionine (Met) deficiency is known to divert folate away from de novo biosyntheses of purines and the pyrimidine, thymidylate, to the resynthesis of Met resulting in deoxynucleoside triphosphate imbalance. We have recently shown that Met can easily be depleted and methylation can be impaired by exposure to a model glutathione (GSH)-depleting agent, bromobenzene (BB). GSH depletion-induced Met depletion, therefore, could cause thymidylate insufficiency for DNA repair synthesis. The administration of thymidine (Thy) should repair this impairment. When this hypothesis was examined in the present study, several interesting results were found. The administration of Thy labeled with [2-14C]Thy to BB-treated Syrian hamsters at either 1, 5, 7 or 9 h after BB resulted in an attenuation of liver toxicity. Intrahepatic hemorrhage, which is a typical characteristic of BB toxicity in the Syrian hamster, was decreased in the BB + Thy groups. The attenuation of liver toxicity was accompanied by a progressive increase of Thy incorporation into liver genomic DNA at 24 h after BB. With respect to the time points chosen for Thy administration, Thy incorporation found in the BB + Thy groups were 2-, 2-, 3- and 4-fold of the controls that received only Thy. The results provide evidence that BB causes a progressive increase of thymidylate insufficiency in liver cells. Thymidylate insufficiency is due to Met depletion, a depletion that occurs as a result of GSH depletion.


The effect of paragigantocellularis lateralis lesion on conditioned place preference (CPP) in presence or absence of alpha2 adrenergic agonist (clonidine) in male rats.
The nucleus paragigantocellularis lateralis (LPGi) is located in the rostral ventrolateral medulla (RVLM), a brain stem region that regulates homeostatic functions such as blood pressure and cardiovascular reflexes, respiration, pain and opiate withdrawal syndrome. LPGi has many anatomical relationships with important nuclei such as arcute nucleus, caudal raphe nucleus, periaqueductal gray (PAG), locus coeruleus (LC), and dentate. In this study we have examined the role of LPGi in the conditioned place preference (CPP) induced by morphine in the presence and absence of clonidine in the rat. We used 49 male N-MRI rats which were divided into 7 groups randomly: 1: Control, 2: Control+saline, 3: sham control, 4: lesion, 5: lesion +0.02 mg/kg clonidine, 6: lesion +0.2 mg/kg clonidine, 7: lesion +2 mg/kg clonidine. Animals were anaesthetized with ketamine (110 mg/kg) and rampune (Xylazine) (3 mg/kg) mixture. In the process of surgery LPGi nucleus has been destroyed bilaterally by DC electrical current (1 mA, 6 second), with stainless steel electrode placed in stereotaxic coordinates of (AP = 11.8, Lat +/- 1.86 and Depth = 10.5). After the recovery period, they were treated with clonidine one hour before the application of Hand's method to induce CPP. We have not found any significant differences between the results of control, control+saline and sham groups in the CPP test but there is a significant increase in the CPP time between sham and LPGi lesion+saline groups (P < 0.019). Clonidine at different doses (0.02, 0.2 and 2 mg/kg) have decreased CPP time in LPGi lesioned group in comparison with lesioned+saline group as well (p < 0.002). In this study we have also demonstrated that clonidine has not any effects on the CPP time in the intact animals. Our results indicate that LPGi lesion induces CPP. It seems that LPGi is involved in drug reinforcements and also LPGi lesion induces sensitivity to alpha2 adrenergic agonist.


Hypoxia induces neurite outgrowth in PC12 cells that is mediated through adenosine A2A receptors.
Development of the nervous system is a complex process, involving coordinated regulation of diverse cellular processes including proliferation, differentiation and synaptogenesis. Disturbances to brain development such as pre- and perinatal hypoxia have been linked to behavioural and late onset of neurological disorders. This study examines the effect of hypoxia on neurite outgrowth in PC12 cells. Hypoxia not only caused a rapid induction of neurite outgrowth, but also synergistically enhanced nerve growth factor (NGF)-induced neurite outgrowth up to 24 h. Transactivation of TrkA receptors was ruled out since the TrkA inhibitor K252a did not block hypoxia-induced neurite outgrowth. Adenosine deaminase prevented hypoxia-induced neurite outgrowth indicating that the effect is mediated by adenosine. Use of the specific adenosine A2A receptor agonist CGS21680 and antagonist 8-3(chlorostyryl)caffeine demonstrated that activation of this receptor is critical for hypoxia-induced neurite outgrowth. Hypoxia-induced neurite outgrowth was blocked by the adenylate cyclase inhibitor, MDL-12,330A, indicating a role for activation of this enzyme in the pathway. Hypoxia was further shown to cause a decrease in growth-associated protein (GAP)-43 levels and a lack of induction of betaIII tubulin, in contrast to NGF treatment which resulted in increased cellular levels of both of these proteins. These findings suggest that hypoxia induces neurite outgrowth in PC12 cells via a pathway distinct from that activated by NGF. Thus, exposure to hypoxia at critical stages of development may contribute to aberrant neurite outgrowth and could be a factor in the pathogenesis of certain delayed developmental neurological disorders.


A comparative study on first stage versus second stage caesarean section on maternal and perinatal outcome.
Caesarean delivery (C/D) can be done in the first or second stage of labor. One fourth of the primary C/D are reported to be performed in the second stage of the labor but are more complicated compared to the ones performed in the first stage. To compare maternal and perinatal outcomes of caesarean delivery (C/D) performed in the second stage of labor compared with the first stage in the Ethiopian setting. An institution based comparative cross-sectional study was conducted in three teaching hospitals in Addis Ababa, Ethiopia. A sample size calculation for double proportion was used, and, for every second stage C/D, the next three consecutive first stage C/D cases were taken as controls till the desired sample size was achieved. mean and standard deviation for continuous and proportion for categorical variables were used for descriptive statistics. T-test for difference in independent mean and chi square test to compare proportions was utilized. Odds ratio with 95% CI was used to measure the strength of association of selected variables. A total of 3238 deliveries were attended in the three teaching hospitals during the study period making the (C/D rate of 30.1%. Three hundred eighty-eight emergency caesarean delivery cases were enrolled using the aforementioned technique with the proportion of 97 (10.9%) second stage and 291 (89.1%) first stage C/D. The most common indications in the first stage were non-reassuring fetal heart rate pattern (NRFHRP) accounting for 110 (37.8%) followed by arrest or protraction disorder of 68 (23.4%), whereas the commonest indication for the second stage C/D was cephalopelvic disproportion 46 (48.5%). Significant difference observed in the mean blood loss between the second stage and first stage C/D, 552 ml vs. 410 ml, (OR 30.13, 95% CI 16.25-55.85). Similarly, the women in the second stage C/D had longer mean hospital stay and mean longer operation time than first stage C/D, 5.34 vs. 6.96 days, (OR 1.72, 95% CI 1.05-2.85), and 31.12 min vs. 37.5, (OR 2.33, 95% CI 1.33-4.07), respectively. Five caesarean hysterectomies were done for postpartum haemorrhage and four cases of extension of incision site were encountered following second stage C/D compared to none in the first stage C/D. Though no maternal death or significant perinatal complications were encountered, women with second stage C/D had significant maternal morbidities than first stage cesarean delivery. Therefore, utmost effort should be made to avoid the procrastination of the decision for C/D to the second stage, but if such compelling situations are encountered, precautions, like involvement of the most senior person and vigilance to minimize the expected complications is warranted.


Visualized gene network reveals the novel target transcripts Sox2 and Pax6 of neuronal development in trans-placental exposure to bisphenol A.
Bisphenol A (BPA) is a ubiquitous endocrine disrupting chemical in our daily life, and its health effect in response to prenatal exposure is still controversial. Early-life BPA exposure may impact brain development and contribute to childhood neurological disorders. The aim of the present study was to investigate molecular target genes of neuronal development in trans-placental exposure to BPA. A meta-analysis of three public microarray datasets was performed to screen for differentially expressed genes (DEGs) in exposure to BPA. The candidate genes of neuronal development were identified from gene ontology analysis in a reconstructed neuronal sub-network, and their gene expressions were determined using real-time PCR in 20 umbilical cord blood samples dichotomized into high and low BPA level groups upon the median 16.8 nM. Among 36 neuronal transcripts sorted from DAVID ontology clusters of 457 DEGs using the analysis of Bioconductor limma package, we found two neuronal genes, sex determining region Y-box 2 (Sox2) and paired box 6 (Pax6), had preferentially down-regulated expression (Bonferroni correction p-value <10(-4) and log2-transformed fold change ≤-1.2) in response to BPA exposure. Fetal cord blood samples had the obviously attenuated gene expression of Sox2 and Pax6 in high BPA group referred to low BPA group. Visualized gene network of Cytoscape analysis showed that Sox2 and Pax6 which were contributed to neural precursor cell proliferation and neuronal differentiation might be down-regulated through sonic hedgehog (Shh), vascular endothelial growth factor A (VEGFA) and Notch signaling. These results indicated that trans-placental BPA exposure down-regulated gene expression of Sox2 and Pax6 potentially underlying the adverse effect on childhood neuronal development.


Retinoblastoma protein enhances the fidelity of chromosome segregation mediated by hsHec1p.
Retinoblastoma protein (Rb) plays important roles in cell cycle progression and cellular differentiation. It may also participate in M phase events, although heretofore only circumstantial evidence has suggested such involvement. Here we show that Rb interacts, through an IxCxE motif and specifically during G(2)/M phase, with hsHec1p, a protein essential for proper chromosome segregation. The interaction between Rb and hsHec1p was reconstituted in a yeast strain in which human hsHEC1 rescues the null mutation of scHEC1. Expression of Rb reduced chromosome segregation errors fivefold in yeast cells sustained by a temperature-sensitive (ts) hshec1-113 allele and enhanced the ability of wild-type hsHec1p to suppress lethality caused by a ts smc1 mutation. The interaction between Hec1p and Smc1p was important for the specific DNA-binding activity of Smc1p. Expression of Rb restored part of the inactivated function of hshec1-113p and thereby increased the DNA-binding activity of Smc1p. Rb thus increased the fidelity of chromosome segregation mediated by hsHec1p in a heterologous yeast system.


Complexity of prefrontal cortical dysfunction in schizophrenia: more than up or down.
Numerous neuroimaging studies have examined the function of the dorsolateral prefrontal cortex in schizophrenia; although abnormalities usually are identified, it is unclear why some studies find too little activation and others too much. The authors' goal was to explore this phenomenon. They used the N-back working memory task and functional magnetic resonance imaging at 3 T to examine a group of 14 patients with schizophrenia and a matched comparison group of 14 healthy subjects. Patients' performance was significantly worse on the two-back working memory task than that of healthy subjects. However, there were areas within the dorsolateral prefrontal cortex of the patients that were more active and areas that were less active than those of the healthy subjects. When the groups were subdivided on the basis of performance on the working memory task into healthy subjects and patients with high or low performance, locales of greater prefrontal activation and locales of less activation were found in the high-performing patients but only locales of underactivation were found in the low-performing patients. These findings suggest that patients with schizophrenia whose performance on the N-back working memory task is similar to that of healthy comparison subjects use greater prefrontal resources but achieve lower accuracy (i.e., inefficiency) and that other patients with schizophrenia fail to sustain the prefrontal network that processes the information, achieving even lower accuracy as a result. These findings add to other evidence that abnormalities of prefrontal cortical function in schizophrenia are not reducible to simply too much or too little activity but, rather, reflect a compromised neural strategy for handling information mediated by the dorsolateral prefrontal cortex.


Investigation of the concentration- and temperature-dependent motion of colloidal nanoparticles.
Although the motion of a single nanoparticle suspended in a fluid can be easily modeled, things get complicated for non-infinitely diluted systems. Coincidentally, these are the systems of interest in relevant fields such as, nanomedicine, microfluidics and miniaturized energy storage devices. Hence, a better understanding of the dynamics of colloidal nanoparticles is utterly needed. Herein, the motion of colloidal suspension of plasmonic nanoparticles (i.e., gold nanoshells) is investigated via laser speckle imaging. The method relies on the analysis of the speckle pattern generated by colloidal suspensions forced to flow at specific velocities. Temperature-dependent measurements corroborated that the dynamics of non-infinitely diluted nanoparticle suspensions are better described through a diffusive model rather than by the equipartition theorem. Under the tested experimental conditions, an average diffusion velocity between 0.37 and 1.57 mm s-1 was found. Most importantly, these values were largely dependent on the nanoparticle concentration. These results are in agreement with previous reports and indicate the existence of long-range interactions between nanoparticles.


In vivo evaluation of stent patency by 64-slice multidetector CT coronary angiography: shall we do it or not?
The diagnostic performance of in-stent restenosis (ISR) by 64-slice multidetector CT coronary angiography (CTCA) has been reported to be influenced by multiple factors. We evaluated individual factors (stent diameter, material and strut thickness) and therefore determined the proper population for follow-up by using this modality. A total of 171 stents were evaluated in 83 consecutive patients with stents imaged with CTCA and conventional coronary angiography. The stent diameter ranged from 2.25 mm to 4.5 mm. 2 models of stainless steel (Taxus Liberte (Boston Scientific, US), 56 stents and Cypher Select (Cordis, US), 34 stents) and 2 models of cobalt alloy (Endeavor (Medtronic, US), 33 stents and Firebird2 (MicroPort, China), 48 stents) were included. By comparing to conventional coronary angiography, the image quality and diagnostic accuracy for ISR were evaluated. The image quality of Taxus, Endeavor and Firebird are markedly better than Cypher in large caliber group (≧3.0 mm) (P < 0.001). Except for Cypher, all other stents with diameter ≧3.0 mm showed excellent diagnostic accuracy (sensitivity 100%, specificity 94.4-96% whereas stents with diameter <3.0 mm had poor diagnostic accuracy (sensitivity 100%, specificity 33.3-70%). Cypher is the stent with thickest strut in our study, and showed reduced image quality and diagnostic accuracy in all stent size, due to large number of unassessable stents. Among 16 binary ISR, 12 lesions were correctly diagnosed by CTCA while the other 4 lesions were unassessable. The main reason for low specificity in small caliber group is the large number of unassessable stents. CTCA has high diagnostic accuracy to identify ISR in selected stents with a diameter of ≧3.0 mm.


Hepatic encephalopathy and Helicobacter pylori: a critical reappraisal.
Hepatic encephalopathy (HE) is a frequent complication of liver cirrhosis, and plasma ammonia plays a pivotal role in its pathogenesis. Ammonia disposal in cirrhotics depend on intricately balanced enzyme and transport systems, involving liver, large and small bowel, muscle, and kidney. Recently, it has been suggested that Helicobacter pylori could contribute to hyperammonemia in cirrhotics, but conflicting data are available in the literature. This is a systematic review of experimental (animals and humans), epidemiological, case-control, and prospective studies, to evaluate the arguments in favor and against the role of H. pylori in HE pathogenesis. Although H. pylori produces ammonia in the stomach, several studies have shown that both basal ammonia levels and HE prevalence did not significantly differ between cirrhotics with and without infection. Moreover, some prospective studies have documented that both blood ammonia levels and mental status in HE cirrhotics are not significantly affected by H. pylori eradication. Even if a small sub-group of cirrhotics with both a high bacterial density and more severe hepatic impairment seems to benefit by bacterial eradication, data indicate that ammonia production in the stomach by H. pylori urease appears to be inadequate to clinically affect ammonia disposal in the majority of cirrhotic patients. Further studies are warranted in this field.


The cell wall sensor Wsc1p is involved in reorganization of actin cytoskeleton in response to hypo-osmotic shock in Saccharomyces cerevisiae.
The cell wall is essential to preserve osmotic integrity of yeast cells. Some phenotypic traits of cell wall mutants suggest that, as a result of a weakening of the cell wall, hypo-osmotic stress-like conditions are created. Consequent expansion of the cell wall and stretching of the plasma membrane trigger a complex response to prevent cell lysis. In this work we examined two conditions that generate a cell wall and membrane stress: one is represented by the cell wall mutant gas1Delta and the other by a hypo-osmotic shock. We examined the actin cytoskeleton and the role of the cell wall sensors Wsc1p and Mid2p in these stress conditions. In the gas1 null mutant cells, which lack a beta(1,3)-glucanosyltransferase activity required for cell wall assembly, a constitutive marked depolarization of actin cytoskeleton was found. In a hypo-osmotic shock wild-type cells showed a transient depolarization of actin cytoskeleton. The percentage of depolarized cells was maximal at 30 min after the shift and then progressively decreased until cells reached a new steady-state condition. The maximal response was proportional to the magnitude of the difference in the external osmolarity before and after the shift within a given range of osmolarities. Loss of Wsc1p specifically delayed the repolarization of the actin cytoskeleton, whereas Wsc1p and Mid2p were essential for the maintenance of cell integrity in gas1Delta cells. The control of actin cytoskeleton is an important element in the context of the compensatory response to cell wall weakening. Wsc1p appears to be an important regulator of the actin network rearrangements in conditions of cell wall expansion and membrane stretching.


MF498 [N-{[4-(5,9-Diethoxy-6-oxo-6,8-dihydro-7H-pyrrolo[3,4-g]quinolin-7-yl)-3-methylbenzyl]sulfonyl}-2-(2-methoxyphenyl)acetamide], a selective E prostanoid receptor 4 antagonist, relieves joint inflammation and pain in rodent models of rheumatoid and osteoarthritis.
Previous evidence has implicated E prostanoid receptor 4 (EP4) in mechanical hyperalgesia induced by subplantar inflammation. However, its role in chronic arthritis remains to be further defined because previous attempts have generated two conflicting lines of evidence, with one showing a marked reduction of arthritis induced by a collagen antibody in mice lacking EP4, but not EP1-EP3, and the other showing no impact of EP4 antagonism on arthritis induced by collagen. Here, we assessed the effect of a novel and selective EP4 antagonist MF498 [N-{[4-(5,9-diethoxy-6-oxo-6,8-dihydro-7H-pyrrolo[3,4-g]quinolin-7-yl)-3-methylbenzyl]sulfonyl}-2-(2-methoxyphenyl)acetamide] on inflammation in adjuvant-induced arthritis (AIA), a rat model for rheumatoid arthritis (RA), and joint pain in a guinea pig model of iodoacetate-induced osteoarthritis (OA). In the AIA model, MF498, but not the antagonist for EP1, MF266-1 [1-(5-{3-[2-(benzyloxy)-5-chlorophenyl]-2-thienyl}pyridin-3-yl)-2,2,2-trifluoroethane-1,1-diol] or EP3 MF266-3 [(2E)-N-[(5-bromo-2-methoxyphenyl)sulfonyl]-3-[5-chloro-2-(2-naphthylmethyl)phenyl]acrylamide], inhibited inflammation, with a similar efficacy as a selective cyclooxygenase 2 (COX-2) inhibitor MF-tricyclic. In addition, MF498 was as effective as an nonsteroidal anti-inflammatory drug, diclofenac, or a selective microsomal prostaglandin E synthase-1 inhibitor, MF63 [2-(6-chloro-1H-phenanthro[9,10-d]imidazol-2-yl)isophthalonitrile], in relieving OA-like pain in guinea pigs. When tested in rat models of gastrointestinal toxicity, the EP4 antagonist was well tolerated, causing no mucosal leakage or erosions. Lastly, we evaluated the renal effect of MF498 in a furosemide-induced diuresis model and demonstrated that the compound displayed a similar renal effect as MF-tricyclic [3-(3,4-difluorophenyl)-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone], reducing furosemide-induced natriuresis by approximately 50%. These results not only suggest that EP4 is the major EP receptor in both RA and OA but also provide a proof of principle to the concept that antagonism of EP4 may be useful for treatment of arthritis.


Characterization and responsiveness of the Madison 109 lung carcinoma to various antitumor agents.
The Madison 109 (M109) tumor was discovered in 1964 in the lung of a BALB/c mouse. This experimental carcinoma is maintained in vivo by sc passage in the right axillary region. When implanted im (5 X 10(5) cells) into the right hind leg of BALB/c mice for testing, the primary progresses with metastases to the lung, spleen, and liver. The metastases to the lung are visible within 3 weeks and result in the death of the host in about 35 days after tumor implant. Implantation of a lung nodule is tumorigenic and lethal. Pyran polymer therapy delayed the appearance of lung metastases, inhibited the growth of the primary tumor, and significantly increased the lifespan of BALB/c mice inoculated with the M109 tumor. No spontaneous regression has been observed and very few "no takes" have occurred in untreated BALB/c mice inoculated with at least 500 M109 cells. Of the 82 agents tested so far, the M109 model has selected active agents such as actinomycin D, adriamycin, daunorubicin, DNA, procarbazine, and pyran polymer. It has not shown sensitivity as tested to several standard therapeutic agents including cytosine arabinoside, BCNU, hydroxyurea, mechlorethamine, melphalan, triethylenemelamine, and vincristine.


Response of unmyelinated (C) polymodal nociceptors to thermal stimuli applied to monkey's face.
1. The response of C polymodal nociceptors to thermal and mechanical stimuli applied to the monkey's face was recorded extracellulary in the trigeminal ganglion in rhesus monkeys anesthetized with sodium pentobarbital. Conduction velocities, determined from electrical stimulation of receptive fields (RFs), were in the range for unmyelinated C fibers (mean=0.82 m/s, n=20; SD=+/-0.17). With two exceptions cutaneous RFs were single spots (median=2 mm2; n=37) and usually were identical for thermal and mechanical stimuli. The median force threshold for the sample of units was 1.2 g (von Frey technique; n = 39; range = 0.07-8.5 g). 2. Discharges to thermal stimuli were investigated with a feedback-controlled contact thermode which permitted temperature changes less than or equal 12.0 degrees C/s. Thermal thresholds ranged from 38 degree to 49 degree C (median=46 degrees C; n=37), and maximum discharge frequencies were obtained in the noxious heat range (45-55 degrees C). For a graded series of 5 s duration stimuli from an adapting temperature of 35 degrees C, the number of impulses increased as a monotonic function of stimulus intensity over the range from threshold temperature to 50-53 degrees C. Many stimulus-response functions were positively accelerated, and linear regression analyses showed that most units examined were best fit by nonlinear functions. 3. The typical pattern of activity to 5 s duration temperature shifts into the noxious heat range was a short accelerating burst of impulses followed by deceleration to a lower rate of discharge prior to termination of the stimulus. The temporal profile of the discharge of impulses was virtually identical at different adapting temperatures. In units tested with 30 s duration stimuli at 2-6 degrees C above threshold, the mean frequency of discharge during the final 25 s was 1.46 impulses/s (n=6; SD=+/-0.89). 4. Application of noxious heat stimuli a few degrees above threshold temperature typically sensitized or enhanced the response of the unit to subsequent application of heat stimuli. The signs of sensitization consisted of a decrease in threshold temperature, increased frequency of discharge, decreased latency to the first impulse, and afterdischarges. Units failed to respond throughout the duration of 30 s stimuli if the final temperature exceeded 50 degrees C. Depressed responses were sometimes produced by application of intense (greater than or equal 55 degrees C) stimuli, presumably as a result of partial inactivation of the receptor. 5. In a correlative analysis, the latency and pattern of discharge in a sample of units were compared with escape responses in two monkeys to temperature shifts into the noxious heat range (49 and 51 degrees C). The analysis revealed that the discharge of C polymodal nociceptors alone cannot account for fast escape responses, but the discharge may contribute to escape responses which occur more than 3.5 s after the onset of stimulation.


The Role of aDNA in Understanding the Coevolutionary Patterns of Human Sexually Transmitted Infections.
Analysis of pathogen genome data sequenced from clinical and historical samples has made it possible to perform phylogenetic analyses of sexually transmitted infections on a global scale, and to estimate the diversity, distribution, and coevolutionary host relationships of these pathogens, providing insights into pathogen emergence and disease prevention. Deep-sequenced pathogen genomes from clinical studies and ancient samples yield estimates of within-host and between-host evolutionary rates and provide data on changes in pathogen genomic stability and evolutionary responses. Here we examine three groups of pathogens transmitted mainly through sexual contact between modern humans to provide insight into ancient human behavior and history with their pathogens. Exploring ancient pathogen genomic divergence and the ancient viral-host parallel evolutionary histories will help us to reconstruct the origin of present-day geographical distribution and diversity of clinical pathogen infections, and will hopefully allow us to foresee possible environmentally induced pathogen evolutionary responses. Lastly, we emphasize that ancient pathogen DNA research should be combined with modern clinical pathogen data, and be equitable and provide advantages for all researchers worldwide, e.g., through shared data.


Factor V Leiden and prothrombin gene G20210A mutation and in vitro fertilization: prospective cohort study.
The influence of thrombophilia on fertility and on IVF outcome is very controversial. The objectives of this study were: (i) to compare the prevalence of Factor V Leiden (FVL) and prothrombin gene G20210A mutation (PGM) in women undergoing IVF to women with spontaneous pregnancy; (ii) to compare the IVF outcomes and the risk of complications in FVL and PGM carrier to non-carrier women. From March 2005 to December 2009, a total of 510 women requiring IVF were recruited in a prospective cohort study. A separate population of 490 nulliparous women who conceived naturally was also evaluated as fertile controls. All women were tested for the presence of FVL and PGM. The prevalence of thrombophilic mutations was the same among women requiring IVF (6.9%) and women with spontaneous pregnancy (6.9%). A total of 480 patients underwent 1105 IVF cycles. There were 30 women carriers (86 IVF cycles) and 450 non-carriers for thrombophilic mutations (1019 IVF cycles). No significant differences in the mean number of oocytes retrieved and the number of good quality embryos transferred were found between the mutation carrier and non-mutation carrier women; likewise the reproductive outcome and the IVF complications were not statistically different between the two groups. The cumulative live birth rate after six IVF cycles was similar in the mutation carrier and non-mutation carrier women. For the mutation carrier women, the optimistic estimate of cumulative live birth rate after six IVF cycles was 60.8% and the conservative estimate was 50.0%. Corresponding rates for the non-mutation carrier women were 56.8 and 36.2%, respectively. The results of this study suggest that FVL and PGM presence in asymptomatic women and in the absence of other risk factors do not influence IVF outcome, or represent risk factors for ovarian hyperstimulation syndrome (OHSS), or favour thrombosis after IVF. Screening for FVL and PGM does not appear to be justified to identify the patients at the risk for IVF failure, and/or for OHSS, and/or for thrombotic complications.


[The blood-brain barrier. II. Physiological data].
In the first part of this review, the morphological characteristics of the interfaces between blood and some compartments of the central nervous system have been described. The blood-brain barrier (BBB) is situated between the blood and the brain extracellular space (ECS) at the endothelial cells of brain capillaries joined by tight junctions. The blood-cerebrospinal fluid (CSF) barrier is mainly situated at the epithelium of the choroid plexuses. In this second part, the general mechanisms and methods of study of the transport of low molecular weight substances across the BBB are reviewed. In addition water and solute transports across the interfaces are described. The third part will deal with the transport of metabolic substrates and of drugs, with the barrier to neurotransmitters and with the physiological alterations of the permeability of the blood-brain barrier. Blood-brain transport is transcellular. According to the nature of the transported molecule it can be either by diffusion through the membranes (lipophilic molecules) or carrier-mediated (hydrophilic molecules such as metabolic substrates). In vivo methods of study have first been experimental. The recent use of short-lived positron-emitting radioisotopes should extent their use to clinical studies. In vitro measurement of transport on isolated capillaries or choroid plexuses give additional information. Water exchange between plasma and the brain is rapid. However, the permeability of the brain capillaries for water is lower than that of other capillaries and of most membranes, but more similar to that of lipid bilayers and of most tight epithelia. The permeability for water of the choroid plexuses which secrete CSF is much higher than that of brain capillaries. CSF and ECS electrolyte composition is under strict homeostatic regulation, leaving it independent of fluctuations in plasma composition. This constant composition is the consequence of the low permeability of brain capillaries to electrolytes as well as that of the presence of a sodium potassium dependent ATPase on the abluminal membrane of capillaries and the apical membrane of choroid plexuses. This enzyme contributes to the extrusion of potassium from the CSF and ECS and allows a large gradient of potassium to be created between ECS and plasma. A low level of potassium in CSF and ECS is in fact needed for normal nerve conduction.


Catabolite repression by glucose 6-phosphate, gluconate and lactose in Escherichia coli.
While catabolite repression by glucose has been studied extensively and is understood in large detail in Enterobacteriaceae, catabolite repression by carbohydrates that are not transported by the phosphotransferase system (PTS) has always remained an enigma. Examples of non-PTS carbohydrates that cause catabolite repression in Escherichia coli are gluconate, lactose and glucose 6-phosphate. In this article it is shown that enzyme IIA(Glc) of the PTS is not involved in catabolite repression by these carbon sources. Carbon sources that caused strong catabolite repression of beta-galactosidase lowered the concentration of both cAMP and the cAMP receptor protein (CRP). A strong correlation was found between the amounts of cAMP and CRP and the strength of the repression. The levels of cAMP and CRP were modulated in various ways. Neither overproduction of CRP nor an increased cAMP concentration could completely relieve the repression by glucose 6-phosphate, lactose and gluconate. Simultaneously increasing the cAMP and the CRP levels was lethal for the cells. In a mutant expressing a constant amount of cAMP-independent CRP* protein, catabolite repression was absent. The same was found in a mutant in which lac transcription is independent of cAMP/CRP. These results, combined with the fact that both the cAMP and the CRP levels are lowered by glucose 6-phosphate, lactose and gluconate, lead to the conclusion that the decreased cAMP and CRP levels are the cause of catabolite repression by these non-PTS carbon sources.


Assessment of some key indicators of the ecological status of an African freshwater lagoon (Lagoon Aghien, Ivory Coast).
The supply of drinking water is a vital challenge for the people who live on the African continent, as this continent is experiencing strong demographic growth and therefore increasing water demands. To meet these needs, surface water resources are becoming increasingly mobilized because underground resources are not always available or have already been overexploited. This situation is the case in the region of Abidjan in the Ivory Coast, where the drinking water deficit is a growing problem and it is therefore necessary to mobilize new water resources to ensure the supply of drinking water. Among the potential resources, local managers have identified a freshwater lagoon, Lagoon Aghien, That is in close proximity to the city of Abidjan. With the aim of enhancing knowledge on the ecological functioning of the lagoon and contributing to the assessment of its ability to provide drinking water, several physical and chemical parameters of the water and the phytoplankton community of the lagoon were monitored for 17 months (December 2016-April 2018) at six sampling stations. Our findings show that the lagoon is eutrophic, as evidenced by the high concentrations of total phosphorus (>140 μg L-1), nitrogen (1.36 mg L-1) and average chlorophyll-a (26 to 167 μg L-1) concentrations. The phytoplankton community in the lagoon is dominated by genera typical of eutrophic environments including mixotrophic genera such as Peridinium and by cyanobacteria such as Cylindrospermopsis/Raphidiopsis, Microcystis and Dolichospermum that can potentially produce cyanotoxins. The two rainfall peaks that occur in June and October appeared to be major events in terms of nutrient flows entering the lagoon, and the dynamics of these flows are complex. Significant differences were also found in the nutrient concentrations and to a lesser extent in the phytoplankton communities among the different stations, especially during the rainfall peaks. Overall, these results reveal that the quality of the lagoon's water is already severely degraded, and this degradation could increase in future years due to increasing urbanization in the watershed. These results therefore raise questions about the potential use of the lagoon as a source of drinking water if measures are not taken very quickly to protect this lagoon from increasing eutrophication and other pollution sources.


Murine endometrial-derived mesenchymal stem cells suppress experimental autoimmune encephalomyelitis depending on indoleamine-2,3-dioxygenase expression.
Cellular therapy with mesenchymal stem cells (MSCs) is a huge challenge for scientists, as little translational relevance has been achieved. However, many studies using MSCs have proved their suppressive and regenerative capacity. Thus, there is still a need for a better understanding of MSCs biology and the establishment of newer protocols, or to test unexplored tissue sources. Here, we demonstrate that murine endometrial-derived MSCs (meMSCs) suppress Experimental Autoimmune Encephalomyelitis (EAE). MSC-treated animals had milder disease, with a significant reduction in Th1 and Th17 lymphocytes in the lymph nodes and in the central nervous system (CNS). This was associated with increased Il27 and Cyp1a1 expression, and presence of IL-10-secreting T CD4+ cells. At EAE peak, animals had reduced CNS infiltrating cells, histopathology and demyelination. qPCR analysis evidenced the down-regulation of several pro-inflammatory genes and up-regulation of indoleamine-2,3-dioxygenase (IDO). Consistently, co-culturing of WT and IDO-/- meMSCs with T CD4+ cells evidenced the necessity of IDO on the suppression of encephalitogenic lymphocytes, and IDO-/- meMSCs were not able to suppress EAE. In addition, WT meMSCs stimulated with IL-17A and IFN-γ increased IDO expression and secretion of kynurenines in vitro, indicating a negative feedback loop. Pathogenic cytokines were increased when CD4+ T cells from AhR-/- mice were co-cultured with WT meMSC. In summary, our research evidences the suppressive activity of the unexplored meMSCs population, and shows the mechanism depends on IDO-kynurenines-Aryl hydrocarbon receptor (AhR) axis. To our knowledge this is the first report evidencing that the therapeutic potential of meMSCs relying on IDO expression.


Is an Empty Nest Best?: Coresidence With Adult Children and Parental Marital Quality Before and After the Great Recession.
Since the Great Recession, the proportion of young adults living with their parents has risen steadily in the United States. Research on coresidence with adult children and parental marital quality is mixed, but marital quality may suffer if children coreside under certain circumstances. When coresidence signifies a deviation from normative expectations, it may be a source of stress in parents' marriages. Further, living with adult children who are suffering problems may be especially detrimental to parental marital quality. Middle-aged parents (N = 287; mean age = 50.65) completed measures of marital quality, child problems, and coresidence at 2 time points, at the onset of the Great Recession in 2008 and again in 2013. Regression analyses estimating marital quality from coresidence status revealed that coresidence with a child was associated with lower parental marital quality in 2008, but not in 2013 (when it may be considered more normative to have adult children living in the home). Additional analyses showed living with a child who was suffering problems was associated with lower marital quality in 2013. These findings suggest that coresidence may be detrimental to marital quality, but perhaps only when coresidence is nonnormative or when coresidence co-occurs with child problems.


Combination therapy with adriamycin and interleukin 2 augments immunity against murine renal cell carcinoma.
The mechanism(s) through which combination treatment with Adriamycin and recombinant interleukin 2 (rIL2) affects murine renal cell carcinoma was investigated. A single dose of Adriamycin (ADM) administered 1 day after implantation of tumor cells s.c. delayed the formation of tumor and significantly extended the median survival time. About 40% of the treated mice remained tumor free for more than 6 months. Treating mice with a similar dose of ADM 12 to 16 days after implantation of tumor caused complete regression of established tumors within 10 to 12 days in more than 90% of the mice. However, after a transient tumor free period of 15 to 20 days recurrence of tumor was observed in all treated mice. In contrast, a regimen that included treating tumor bearing mice with a single dose of ADM followed by a daily i.p. injection of rIL2 5 x 10(4) units for 10 days delayed the recurrence of tumors and significantly prolonged the survival time compared to the median survival time of mice treated with ADM alone. About 20 to 30% tumor bearing mice remained tumor free for 4 months following treatment with ADM and rIL2. Treatment with rIL2 alone produced no antitumor response. In addition, rIL2 itself was not inhibitory for tumor cell growth nor did it modulate the cytotoxic response of renal cell carcinoma cells to ADM in vitro. Mice that were cured following treatment with ADM and rIL2 were resistant to a rechallenge with viable tumor cells, and cured mice also expressed a tumor specific T-cell mediated delayed hypersensitivity reaction. The immune cells that mediate tumor rejection were identified as Thy-1.2+ T-cells. Taken together, these results indicate that antitumor activity of combined Adriamycin/rIL2 treatment is at least partly attributable to the production of tumor specific immunity.


[The specific characteristics of the vertebrate electroretinogram evoked by x-rays].
Summarised electroretinograms (ERGs) of dark-adaptable retina of Rana temporaria were studied in situ with an automated experimental device with programmable control and first and second derivatives electron differentiation. X-rays were shown to induce the ERG composed of two components which were conventionally designated as the first and second roentgen responses (R-1 and R-2). These differ by their parameters from the ERGs induced by red and blue light in the same experiment. Administration of sodium azide, sodium nitrite, monoiodine acetate and other compounds changed the waves of R-1 and R-2 as well as photo-induced ERGs. This suggests their relative independence and different mechanism of initiation, and allows their partial isolation for separate studying.


An outbreak of Salmonella Typhimurium phage type 64 gastroenteritis linked to catered luncheons in Adelaide, South Australia, June 2005.
Salmonella sp. are important causes of foodborne illness, with restaurants and catered functions being commonly reported settings for outbreaks. In June 2005 an investigation commenced following reports of gastrointestinal illness in attendees at luncheons catered by an Adelaide café, as well as persons eating at the café itself. The investigation sought to determine the existence of an outbreak, identify a source and method of transmission and implement public health measures to prevent further cases. Lists of luncheon attendees were obtained from function organisers. A retrospective cohort study was commenced using a structured questionnaire developed from the café's menu listings. A suspected case was defined as a person developing two or more gastrointestinal symptoms after attending a luncheon catered by the café. A case series investigation was used for café diners. Of the 102 respondents, 61 (60%) met the case definition with 32 subsequently confirmed as Salmonella Typhimurium phage type 64 (STM 64) infections. Of the 61 cases, 59 (96%) reported eating a bread roll. STM 64 was detected in raw defrosted chicken recovered from the café's kitchen. This suggested cross-contamination from the chicken to one or more ingredients common to the bread rolls was the route of infection. To prevent further cases, perishable goods were discarded, the café was closed, the premises cleaned, then restrictions were placed on the types of foods served. This investigation's findings highlight the importance of safe food handling and hand hygiene in commercial food preparation.


Unravelling sleep problems in treated and untreated children with ADHD.
This study investigated parental perception of sleep problems in stimulant treated and untreated children with Attention Deficit Hyperactivity Disorder (ADHD). Parents of 135 psychiatric clinic referred children and 83 pediatric outpatients completed a sleep questionnaire and the Child Behavior Checklist. Moderate to severe "sleep problems" reportedly occurred at least once a week in 19.3% of children with ADHD, 13.3% of the psychiatric controls, and 6.2% of the pediatric controls. Children with ADHD treated with stimulants were reported to display a higher prevalence of nightly "severe" sleep problems than did untreated children with ADHD. Almost a third (29%) of stimulant treated ADHD children were reported to display increased sleep latency or insomnia every night versus 10% of untreated children with ADHD. Despite the high prevalence of sleep related problems in ADHD, the significance of the association between delayed sleep onset and ADHD with regard to etiology and management of ADHD is still poorly understood.


A general strategy for printing colloidal nanomaterials into one-dimensional micro/nanolines.
Though patterned one-dimensional (1D) micro/nanoline arrays are of great importance in the field of integrated circuits and optoelectronics, the fabrication of high-precision micro/nanolines with excellent optical and electrical performance remains a great challenge. Herein, a general strategy for printing 1D micro/nanolines is proposed by manipulating the self-assembly of functional nanoparticles as a multilayer or monolayer stack with a single-nanoparticle width. This method is universal for dispersible nanoparticles, and the silver nanoparticle was selected as a model nanoparticle due to its good conductivity, dispersibility and narrow-size distribution. The results indicate that the morphologies of printed micro/nanolines can be precisely regulated by the substrate wettability and the suspension concentration. Specifically, 1D nanoparticle-assembled architectures are printed as a monolayer stack on the substrate with a low contact angle (below 45°), while a multilayer stack is formed on the substrate with a high contact angle (above 50°) or a high concentration (more than 0.12%). The controllability of micro/nanoline morphologies can be interpreted through the influence of the three phase contact line slipping motion and the nanoparticle diffusion on diverse substrates at different concentrations. Alteration of the printing template structures enables the intervals of 1D micro/nanolines to span from 16 μm to 48 μm. These results provide an efficient methodology for fabricating micro/nano-circuits or optics and strengthening the understanding of the self-assembling process.


Targeting Mosquitoes through Generation of an Insecticidal RNAi Yeast Strain Using Cas-CLOVER and Super PiggyBac Engineering in Saccharomyces cerevisiae.
The global deployment of RNAi yeast insecticides involves transitioning from the use of laboratory yeast strains to more robust strains that are suitable for scaled fermentation. In this investigation, the RNA-guided Cas-CLOVER system was used in combination with Piggybac transposase to produce robust Saccharomyces cerevisiae strains with multiple integrated copies of the Sh.463 short hairpin RNA (shRNA) insecticide expression cassette. This enabled the constitutive high-level expression of an insecticidal shRNA corresponding to a target sequence that is conserved in mosquito Shaker genes, but which is not found in non-target organisms. Top-expressing Cas-CLOVER strains performed well in insecticide trials conducted on Aedes, Culex, and Anopheles larvae and adult mosquitoes, which died following consumption of the yeast. Scaled fermentation facilitated the kilogram-scale production of the yeast, which was subsequently heat-killed and dried. These studies indicate that RNAi yeast insecticide production can be scaled, an advancement that may one day facilitate the global distribution of this new mosquito control intervention.


MUC1 peptide epitopes associated with five different H-2 class I molecules.
We have previously described the induction of murine CD8+ major histocompatibility complex (MHC) class I-restricted cytotoxic T cells (CTL) recognizing the 20-amino acid repeat region of the human mucin 1 (MUC1) variable number of tandem repeats region (VNTR), a mucin greatly increased in expression in breast cancer and proposed as a target for immunotherapy. In that study, CTL could detect MUC1 peptides associated with the MHC of all nine strains examined, and we now report the different epitopes presented by five different MHC class I molecules. The epitopes were defined in CTL assays using peptide-pulsed phytohemagglutinin blasts or MHC class I-transfected L cells as targets; in addition, peptide binding assays and T cell proliferation studies were performed. Within the 20-amino acid VNTR, nine potential epitopes could be defined. The epitopes for the four MHC class I molecules [Kb (three epitopes), Dd, Ld and Kk] were closely related, all containing the amino acids PDTRPAP. For Db, three epitopes were identified, all containing APGSTAP. Most of the epitopes did not contain a consensus motif for the particular MHC class I allele, and bound with low 'affinity', compared with known high-affinity peptides. CD8+ T cell proliferation also occurred to the same MHC class I-presented epitopes. Finally, when conventional anchor residues were introduced into the peptides, peptide binding increased, whereas CTL recognition was either retained (Kb) or lost (Db) depending on the epitope.


Advancements in Improving Selectivity of Metal Oxide Semiconductor Gas Sensors Opening New Perspectives for Their Application in Food Industry.
Volatile compounds not only contribute to the distinct flavors and aromas found in foods and beverages, but can also serve as indicators for spoilage, contamination, or the presence of potentially harmful substances. As the odor of food raw materials and products carries valuable information about their state, gas sensors play a pivotal role in ensuring food safety and quality at various stages of its production and distribution. Among gas detection devices that are widely used in the food industry, metal oxide semiconductor (MOS) gas sensors are of the greatest importance. Ongoing research and development efforts have led to significant improvements in their performance, rendering them immensely useful tools for monitoring and ensuring food product quality; however, aspects related to their limited selectivity still remain a challenge. This review explores various strategies and technologies that have been employed to enhance the selectivity of MOS gas sensors, encompassing the innovative sensor designs, integration of advanced materials, and improvement of measurement methodology and pattern recognize algorithms. The discussed advances in MOS gas sensors, such as reducing cross-sensitivity to interfering gases, improving detection limits, and providing more accurate assessment of volatile organic compounds (VOCs) could lead to further expansion of their applications in a variety of areas, including food processing and storage, ultimately benefiting both industry and consumers.


Cellular physiology of the turtle visual cortex: synaptic properties and intrinsic circuitry.
We have examined the synaptic physiology of the isolated dorsal cortex of the turtle, Pseudemys scripta elegans. Electrical stimulation of afferent pathways elicited distinct, stereotyped responses in pyramidal and stellate neurons. Single shocks evoked a long-lasting barrage of excitatory postsynaptic potentials (EPSPs) in stellate cells, and led to a burst of several action potentials. Under the same circumstances, pyramidal cells displayed a small amount of short-latency excitation, but this was accompanied by a profound and prolonged set of inhibitory post-synaptic potentials (IPSPs). Synaptic excitation of the distal dendrites of pyramidal cells could evoke dendritic action potentials that were visible at the soma as small all-or-none spikes rising from the hyperpolarized level of the IPSP. There appeared to be two mechanistically different types of IPSPs in pyramidal cells. The first occurred at short latency, could produce a very large conductance increase, reversed polarity at -71 mV, and was chloride-dependent. The second was generally smaller and more protracted, had a relatively negative reversal potential of -85 to -95 mV, and was insensitive to chloride injection. Focal application of small doses of the putative inhibitory neurotransmitter gamma-aminobutyric acid (GABA) onto the somata of pyramidal cells caused a conductance increase and hyperpolarization. This response had features in common with the short-latency IPSP, including an identical reversal potential. Application of large doses of GABA to the somata of pyramidal cells or smaller doses to their dendrites elicited multiphasic or purely depolarizing responses that were at least partly due to time- or space-dependent shifts of the equilibrium potential of the response. Bicuculline methiodide, a potent GABA antagonist, depressed both the responses to GABA and the short-latency IPSP, but not the long-latency IPSP; synchronized epileptiform burst discharges also resulted. These findings, together with responses to locally applied electric shocks and the excitatory amino acid glutamate, suggested that inhibition of pyramidal cells was generated intrinsically by stellate cells, and that the cortical circuit provides pathways for both feedforward and feedback GABAergic inhibition. The data also suggest that pyramidal cells are mutually excitatory. These features are similar to the basic intrinsic circuitry in the telencephalic cortices of mammals.


Characterization of phagolysosomal simulant fluid for study of beryllium aerosol particle dissolution.
A simulant of phagolysosomal fluid is needed for beryllium particle dissolution research because intraphagolysosomal dissolution is believed to be a necessary step in the cellular immune response associated with development of chronic beryllium disease. Thus, we refined and characterized a potassium hydrogen phthalate (KHP) buffered solution with pH 4.55, termed phagolysosomal simulant fluid (PSF), for use in a static dissolution technique. To characterize the simulant, beryllium dissolution in PSF was compared to dissolution in the J774A.1 murine cell line. The effects of ionic composition, buffer strength, and the presence of the antifungal agent alkylbenzyldimethylammonium chloride (ABDC) on beryllium dissolution in PSF were evaluated. Beryllium dissolution in PSF was not different from dissolution in the J774A.1 murine cell line (p = 0.78) or from dissolution in another simulant having the same pH but different ionic composition (p = 0.73). A buffer concentration of 0.01-M KHP did not appear adequate to maintain pH under all conditions. There was no difference between dissolution in PSF with 0.01-M KHP and 0.02-M KHP (p = 0.12). At 0.04-M KHP, beryllium dissolution was increased relative to 0.02-M KHP (p = 0.02). Use of a 0.02-M KHP buffer concentration in the standard formulation for PSF provided stability in pH without alteration of the dissolution rate. The presence of ABDC did not influence beryllium dissolution in PSF (p = 0.35). PSF appears to be a useful and appropriate model of in vitro beryllium dissolution when using a static dissolution technique. In addition, the critical approach used to evaluate and adjust the composition of PSF may serve as a framework for characterizing PSF to study dissolution of other metal and oxide particles.


[Market-based medicine or patient-based medicine?].
The health care has evolved over the centuries from a theocentric model to a model centered on man, environment and society. The process of neoliberal globalization has changed the relationship between the components of the health system and population. The active participation of organizations such as the World Trade Organization, the International Monetary Fund and the World Bank by the techno-medical industrial complex tends to make the health care in a model focused on economy. This, impacts negatively on all components in the process of health care and have an adverse effect on the humanized care. The analysis of each sector in particular and their interactions shows the effects of this change. Alternatives are proposed for each sector to contribute to a model of care focused on the patient, their family and the social environment.


[Relationship between MTA1 expression and invasive and metastatic ability of cervical cancer cell].
To investigate the relationship between metastasis-associated gene 1 (MTA1) expression and invasive and metastatic ability of cervical cancer cell. Three kinds of plasmids pcDNA3 (control group), pcDNA3-MTA1 (MTA1 group) and pSilencer3.1-MTA1-siRNA (MTA1-siRNA group) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability. Compared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P < 0.05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ± 6, 169 ± 10 and 57 ± 5, respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group (P < 0.05). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69.3 ± 3.6)%, (80.4 ± 5.6)% and (39.2 ± 7.4)% separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P < 0.05). In the migration assay, the number of cells migrated to the bottom side of the membrane in control, MTA1 and MTA1-siRNA group were 153 ± 17, 247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19, 354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA1-siRNA group showed lower cell migration and invasion ability (P < 0.05). In cell cycle experiment, no significant differences of cell proportions including G(1), S and G(2) stage were found among three groups (P > 0.05). In animal experiment, compared with control group, MTA1 group showed accelerated tumor formation and growth, while the MTA1-siRNA group showed the reverse effect (P < 0.05). MTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.