Chronic activation of ORL1 receptor induces supersensitization of adenylyl cyclase.
The opioid receptor-like (ORL1) receptor regulates a variety of effectors shared by its close relatives, the opioid receptors. Supersensitization of adenylyl cyclase (AC) is a hallmark of cellular tolerance induced by chronic activation of opioid receptors. To examine if chronic activation of the ORL1 receptor leads to a similar adaptation, a HEK293 cell line stably expressing the human ORL1 receptor (293/ORL1) was established. Chronic treatment of 293/ORL1 cells with nociceptin/OFQ resulted in enhanced AC activity in response to forskolin stimulation. The AC supersensitivity was blocked by pertussis toxin, indicating the involvement of Gi/Go proteins. Likewise, chronic activation of endogenous ORL1 receptors in the neuroblastoma SK-N-SH cells led to Gi/Go-mediated AC supersensitization.


Evaluation of Chlorine Treatment Levels for Inactivation of Human Norovirus and MS2 Bacteriophage during Sewage Treatment.
This study examined the inactivation of human norovirus (HuNoV) GI.1 and GII.4 by chlorine under conditions mimicking sewage treatment. Using a porcine gastric mucin-magnetic bead (PGM-MB) assay, no statistically significant loss in HuNoV binding (inactivation) was observed for secondary effluent treatments of ≤25 ppm total chlorine; for both strains, 50 and 100 ppm treatments resulted in ≤0.8-log10 unit and ≥3.9-log10 unit reductions, respectively. Treatments of 10, 25, 50, and 100 ppm chlorine inactivated 0.31, 1.35, >5, and >5 log10 units, respectively, of the norovirus indicator MS2 bacteriophage. Evaluation of treatment time indicated that the vast majority of MS2 and HuNoV inactivation occurred in the first 5 min for 0.2-μm-filtered, prechlorinated secondary effluent. Free chlorine measurements of secondary effluent seeded with MS2 and HuNoV demonstrated substantial oxidative burdens. With 25, 50, and 100 ppm treatments, free chlorine levels after 5 min of exposure ranged from 0.21 to 0.58 ppm, from 0.28 to 16.7 ppm, and from 11.6 to 53 ppm, respectively. At chlorine treatment levels of >50 ppm, statistically significant differences were observed between reductions for PGM-MB-bound HuNoV (potentially infectious) particles and those for unbound (noninfectious) HuNoV particles or total norovirus particles. While results suggested that MS2 and HuNoV (measured as PGM-MB binding) behave similarly, although not identically, both have limited susceptibility to chlorine treatments of ≤25 ppm total chlorine. Since sewage treatment is performed at ≤25 ppm total chlorine, targeting free chlorine levels of 0.5 to 1.0 ppm, these results suggest that traditional chlorine-based sewage treatment does not inactivate HuNoV efficiently.IMPORTANCE HuNoV is ubiquitous in sewage. A receptor binding assay was used to assess inactivation of HuNoV by chlorine-based sewage treatment, given that the virus cannot be routinely propagated in vitro Results reported here indicate that chlorine treatment of sewage is not effective for inactivating HuNoV unless chlorine levels are above those routinely used for sewage treatment.


[Isoelectric characterization of extracellular polygalacturanases of various Aspergillus alliaceus strains].
The composition of pectin hydrolase complexes produced by various Aspergillus alliaceus strains was studied under the conditions of induction, catabolite repression, or constitutive synthesis. The strains were found similar in terms of the polygalacturonase spectrum and different with regard to the levels of endo- and exoenzyme activities. The analysis of the zymograms of inducible polygalacturonases revealed that all tested cultures contained at least 24 molecular forms of polygalacturonase. Taking into account only the three molecular forms typical of all analyzed strains of A. alliaceus with pI values of 5.7, 5.9, and 6.3, one can use the spectrum of constitutive, catabolite repression-resistant polygalacturonases as an additional taxonomic species criterion.


Modeling Infrared Signal Reflections to Characterize Indoor Multipath Propagation.
In this paper, we propose a model to characterize Infrared (IR) signal reflections on any kind of surface material, together with a simplified procedure to compute the model parameters. The model works within the framework of Local Positioning Systems (LPS) based on IR signals (IR-LPS) to evaluate the behavior of transmitted signal Multipaths (MP), which are the main cause of error in IR-LPS, and makes several contributions to mitigation methods. Current methods are based on physics, optics, geometry and empirical methods, but these do not meet our requirements because of the need to apply several different restrictions and employ complex tools. We propose a simplified model based on only two reflection components, together with a method for determining the model parameters based on 12 empirical measurements that are easily performed in the real environment where the IR-LPS is being applied. Our experimental results show that the model provides a comprehensive solution to the real behavior of IR MP, yielding small errors when comparing real and modeled data (the mean error ranges from 1% to 4% depending on the environment surface materials). Other state-of-the-art methods yielded mean errors ranging from 15% to 40% in test measurements.


One case of sublingual gland mucosa-associated lymphoid tissue lymphoma.
Mucosa-associated lymphoid tissue (MALT) lymphoma is a type of marginal zone B-cell lymphoma that occurs outside the lymph nodes in mucosal tissue. It accounts for 6-8 % of non-Hodgkin's lymphomas. MALT lymphoma of the salivary gland is a rare disease, with primary tumors in the salivary gland accounting for 2-5 % of salivary gland tumors. The most common site is the parotid gland (80 %), followed by the submandibular gland (14 %), minor salivary glands, and sublingual gland (5 %). Patients with salivary gland MALT lymphoma often have autoimmune diseases such as Sjogren's syndrome and rheumatoid arthritis. Primary malignant tumors of the sublingual gland account for less than 1 % of cases, and preoperative diagnosis is difficult, often requiring biopsy for confirmation. To our knowledge, there are no reports of MALT lymphoma arising from the sublingual gland. We report a case of MALT lymphoma originating from the sublingual gland in a patient with a history of hypertension, diabetes, cerebral infarction, and non-traumatic numbness of the right lower limb.


Stimulation of albumin endocytosis by cationized ferritin in cultured aortic smooth muscle cells.
Anionic microdomains within the aortic smooth muscle cell (SMC) surface glycocalyx represent a potential barrier to the endocytosis of anionic plasma proteins. Cultured SMCs exposed briefly to cationized ferritin (CF) exhibit ultrastructural aggregations of surface anionic sites resulting in intervening areas essentially devoid of anionic charge. Preincubation of cultured aortic medial SMCs with 0.2 mg/ml CF for 1 minute at 37 C resulted in a 4-fold increase in binding and a 13-fold increase in internalization of 125I-human serum albumin (125I-HSA) relative to cells pretreated with native ferritin. When both the CF preincubation and the endocytosis were performed at 4 C, the influence of CF was abolished. Studies at 4 C indicated that CF pretreatment of SMC at 37 C induced high affinity (Kd = 1.5 nM) saturable 125I-HSA binding, in addition to low-affinity nonsaturable binding. These results were further confirmed by binding competition studies using increasing concentrations of unlabeled HSA. In contrast, low-density lipoprotein, a large anionic molecule, failed to compete with 125I-HSA for binding sites on CF-pretreated SMCs at either 4 or 37 C. Pulse-chase studies at 37 C indicated that 20-30% of internalized 125I-HSA was degraded, and 40-50% exocytosed within 24 hours in CF-treated cells. CF pretreatment of the SMCs did not significantly enhance the uptake of 14C-sucrose as a measure of fluid-phase endocytosis at 30 and 60 minutes. The results of these studies emphasize the potentially important regulatory roles of cell-surface anionic charge distribution and cationic molecules in cellular endocytosis.


A Personalized Cancer Nanovaccine that Enhances T-Cell Responses and Efficacy Through Dual Interactions with Dendritic Cells and T Cells.
Conventional approaches to developing therapeutic cancer vaccines that primarily activate tumor-specific T cells via dendritic cells (DCs) often demonstrate limited efficacy due to the suboptimal activation of these T cells. To address this limitation, here a therapeutic cancer nanovaccine is developed that enhances T cell responses by interacting with both DCs and T cells. The nanovaccine is based on a cancer cell membrane nanoparticle (CCM-MPLA) that utilizes monophosphoryl lipid A (MPLA) as an adjuvant. To allow direct interaction between the nanovaccine and tumor-specific T cells, anti-CD28 antibodies (aCD28) are conjugated onto CCM-MPLA, resulting in CCM-MPLA-aCD28. This nanovaccine activates tumor-specific CD8+ T cells in both the presence and absence of DCs. Compared with nanovaccines that interact with either DCs (CCM-MPLA) or T cells (CCM-aCD28), CCM-MPLA-aCD28 induces more potent responses of tumor-specific CD8+ T cells and exhibits a higher antitumor efficacy in tumor-bearing mice. No differences in T cell activation efficiency and therapeutic efficacy are observed between CCM-MPLA and CCM-aCD28. This approach may lead to the development of effective personalized therapeutic cancer vaccines prepared from autologous cancer cells.


Validation of the EuroClonality-NGS DNA capture panel as an integrated genomic tool for lymphoproliferative disorders.
Current diagnostic standards for lymphoproliferative disorders include multiple tests for detection of clonal immunoglobulin (IG) and/or T-cell receptor (TCR) rearrangements, translocations, copy-number alterations (CNAs), and somatic mutations. The EuroClonality-NGS DNA Capture (EuroClonality-NDC) assay was designed as an integrated tool to characterize these alterations by capturing IGH switch regions along with variable, diversity, and joining genes of all IG and TCR loci in addition to clinically relevant genes for CNA and mutation analysis. Diagnostic performance against standard-of-care clinical testing was assessed in a cohort of 280 B- and T-cell malignancies from 10 European laboratories, including 88 formalin-fixed paraffin-embedded samples and 21 reactive lesions. DNA samples were subjected to the EuroClonality-NDC protocol in 7 EuroClonality-NGS laboratories and analyzed using a bespoke bioinformatic pipeline. The EuroClonality-NDC assay detected B-cell clonality in 191 (97%) of 197 B-cell malignancies and T-cell clonality in 71 (97%) of 73 T-cell malignancies. Limit of detection (LOD) for IG/TCR rearrangements was established at 5% using cell line blends. Chromosomal translocations were detected in 145 (95%) of 152 cases known to be positive. CNAs were validated for immunogenetic and oncogenetic regions, highlighting their novel role in confirming clonality in somatically hypermutated cases. Single-nucleotide variant LOD was determined as 4% allele frequency, and an orthogonal validation using 32 samples resulted in 98% concordance. The EuroClonality-NDC assay is a robust tool providing a single end-to-end workflow for simultaneous detection of B- and T-cell clonality, translocations, CNAs, and sequence variants.


[Evolution of granulomas induced by BCG vaccine administered in the experiment].
The time course of hepatic granulomatosis was studied on a CBAxC57B1/6 murine model of a vaccinal process induced by BCG vaccine. It was found that over the natural course of inflammation, there were changes in the spectrum of granulomas from epithelioid cellular to macrophageal and fibrotic ones with stabilization of their sizes, averaged cellular composition and density in the organ. Long-term use of isoniazid caused a considerably (2-fold) reduction in the number of granulomas in the organ and thus a decrease in the level of its fibrotic process. Presumably, in the treated animals, the mechanism responsible for a reduction in the number of granulomas is the "dissociation" of macrophageal cells from them.


Pneumococcal serotype distribution among meningitis cases from Togo and Burkina Faso during 2007-2009.
We evaluated pneumococcal serotype/group distribution using polymerase chain reaction (PCR) testing on cerebrospinal fluid collected from patients from Burkina Faso and Togo who presented for care during 2007-2009. We identified 282 pneumococcal meningitis cases based on PCR, latex agglutination, or culture, of which 206 underwent serotyping. Serotype 1 was identified for 18% of serotyped cases from patients aged <5 years and 66% of those aged ≥5 years. The 13-valent and 10-valent pneumococcal conjugate vaccines (PCV-13 and PCV-10) contain 53% of serotypes identified among children age <5 years and 76-77% among persons aged ≥5 years. Pneumococcal meningitis was highly seasonal regardless of serotype. Data from this study emphasize the potential usefulness of PCVs among older children and adults.


Expression of the G-CSF receptor on hematopoietic progenitor cells is not required for their mobilization by G-CSF.
The mechanisms that regulate hematopoietic progenitor cell (HPC) mobilization from the bone marrow to blood have not yet been defined. HPC mobilization by granulocyte colony-stimulating factor (G-CSF), cyclophosphamide (CY), or interleukin-8 but not flt-3 ligand is markedly impaired in G-CSF receptor-deficient (G-CSFR-deficient) mice. G-CSFR is expressed on mature hematopoietic cells, HPCs, and stromal cells, which suggests that G-CSFR signals in one or more of these cell types was required for mobilization by these agents. To define the cell type(s) responsible for G-CSF-dependent mobilization, a series of chimeric mice were generated using bone marrow transplantation. Mobilization studies in these chimeras demonstrated that expression of the G-CSFR on transplantable hematopoietic cells but not stromal cells is required for CY- or G-CSF-induced mobilization. Moreover, in irradiated mice reconstituted with both wild type and G-CSFR-deficient bone marrow cells, treatment with CY or G-CSF resulted in the equal mobilization of both types of HPCs. This result held true for a broad spectrum of HPCs including colony-forming cells, CD34(+) lineage(-) and Sca(+) lineage(-) cells, and long-term culture initiating cells. Collectively, these data provide the first definitive evidence that expression of the G-CSFR on HPCs is not required for their mobilization by G-CSF and suggest a model in which G-CSFR-dependent signals act in trans to mobilize HPCs from the bone marrow.


Cytoplasmic pH regulation in phorbol ester-activated human neutrophils.
Activation of neutrophils by 12-O-tetradecanoylphorbol-13-acetate (TPA) is accompanied by an initial cytoplasmic acidification, followed by an alkalinizing phase due to Na+-H+ countertransport. The source of the acidification, which is fully expressed by activation with TPA in Na+-free or amiloride-containing media, was investigated. The acidification phase was detected also in degranulated and enucleated cytoplasts, ruling out a major contribution by the nucleus or secretory vesicles. Cytoplasmic acidification was found to be associated with an extracellular acidification, suggesting metabolic generation of H+. Two principal metabolic pathways are stimulated in activated neutrophils: the reduction of O2 by NADPH-oxidase and the hexose monophosphate shunt. A good correlation was found between the activity of these pathways and the changes in cytoplasmic pH. Inhibition of superoxide synthesis prevented the TPA-induced cytoplasmic acidification. Moreover, activation of the hexose monophosphate shunt with permeable NADPH-oxidizing agents (in the absence of TPA) also produced a cytoplasmic acidification. Cytoplasmic acidification was also elicited by exogenous diacylglycerol and by other beta-phorbol diesters, which are activators of the kinase, but not by unesterified phorbol or by alpha-phorbol diesters, which are biologically inactive. The results suggest that the cytoplasmic acidification induced by phorbol esters in neutrophils reflects accumulation of H+ liberated during the metabolic burst that follows activation.


[The wound drainage after operation on thyroid gland].
Efficacy of active draining, promoting the most rapid reconvalescence and rehabilitation of the patients, in comparison to passive wound draining was studied up in various diseases of thyroid gland and operative intervention volume. The autors have had concluded, that suturing of operative wound without draining after hemithyroidectomy conduction is possible, as well as a thyroidectomy for nodular and multinodular goiter, if the excised volume of the lobe/gland do not exceed 15 cm3 and in nonapplication of anticoagulants.


Challenges for principles of need in health care.
What challenges must a principle of need for prioritisations in health care meet in order to be plausible and practically useful? Some progress in answering this question has recently been made by Hope, Østerdal and Hasman. This article continue their work by suggesting that the characteristic feature of principles of needs is that they are sufficientarian, saying that we have a right to a minimally acceptable or good life or health, but nothing more. Accordingly, principles of needs must answer two distributive questions: when do we have sufficient and how should we prioritise among those who do not yet have a sufficiency? Furthermore, it is argued that Roger Crisp's theory of need, which combines sufficientarianism with prioritarianism below the threshold of need, is better equipped than alternatives to answer these questions as well as meeting the challenges formulated by Hope, Østerdal and Hasman. However, Crisp's theory faces two major challenges. First, it has to say something about the currency of distribution: a principle of need must be complemented either with a theory on the human good or a theory about the proper goals of health care. Second, it has to say something about where the threshold should be set. However, any attempt to set a threshold seems morally arbitrary in the light of the sufficientarian idea that those just above the threshold never should be given priority over those just below the threshold.


[Geriatric emergencies versus adult emergencies: retrospective analysis of medical emergencies at a general hospital].
The aim of the study was to evaluate the process of the attention to emergencies in patients older than 65 years and to compare it with the same process in adult patients. To this end, 965 clinical records of medical emergencies from the Hospital Central de la Cruz Roja in Madrid were retrospectively evaluated, and data were obtained regarding age, the cause for consultation, the investigations performed and their yield, the administration of drug therapy, the major diagnosis at the time of discharge from the service and the clinical course. It was found that all evaluated diagnostic investigations were carried out with equal or higher frequency in patients older than 65 years and that their mean clinical effectiveness was also higher. In addition, it was found that the patients older than 65 years were more commonly admitted to the hospital through the emergency service than the rest of the population. It was concluded, therefore, that the process of attention to emergencies has differential characteristics in the elderly population, and that if the number and proportion of old people increase as it will presumably happen during the two next decades, the cost of attention to emergencies and the number of emergency hospital admissions will also increase.


Chemiluminescent Analysis of Oxidative Metabolism in Rat Blood under the Influence of Argon and Helium.
We studied the nature of the action of course treatment with argon and helium (1 min, 3 procedures) on the oxidative metabolism in rat blood plasma. The study was performed on 30 Wistar rats divided into 3 groups (n=10 in each group): intact and 2 experimental (treatment of the skin of the back with a stream of argon and helium, respectively). After completion of the treatment course, the intensity of free radical processes, the total antioxidant activity, and malondialdehyde concentration were evaluated in the blood plasma. It was found that argon and helium gas flows provide stimulation of antioxidant systems, but the mechanisms of their effect were different. Treatment with helium did not affect the intensity of free radical processes, but significantly increased the overall antioxidant activity of blood plasma and reduced malondialdehyde concentration in comparison with the effect of argon flow.


Intraoperative floppy iris syndrome: pathophysiology, prevention, and treatment.
To extend upon previous reports, observations, and discussions of intraoperative floppy iris syndrome (IFIS) with the goal of providing new insight into the syndrome's pathophysiology, prevention, and treatment. Following a review of IFIS and its relationship to autonomic pharmacology, evidence for anatomic changes following exposure of humans and other animals to autonomic drugs is described. The clinical implications for these findings are discussed as they relate to the treatment and prevention of this syndrome. IFIS has been associated with the use of adrenergic antagonists even after they have been discontinued years prior to surgery. Some investigators believe that this persistence of IFIS reflects anatomic structural change. Evidence from laboratory experiments and human clinical studies using topically applied and systemic autonomic drugs supports the possibility of anatomic changes coexisting with IFIS observed during cataract surgery. IFIS is a relatively rare syndrome, often associated with the use of systemic alpha-blockers and conditions that influence dilator muscle tone. Laboratory and clinical evidence supports the possibility of anatomic changes following the use of autonomic drugs. The persistence of IFIS years after cessation of treatment with alpha-blockers suggests that the potential risks of discontinuing these drugs prior to cataract surgery outweigh potential benefits.


Segregation and mapping analysis of polymorphic HLA class I restriction fragments: detection of a novel fragment.
An HLA-B7 complementary DNA clone was used as a hybridization probe to analyze the segregation pattern of polymorphic class I restriction fragments in several families whose HLA types had been determined by serological techniques. In one family in which a crossover in the HLA region had occurred, a specific genomic fragment was mapped with respect to the crossover. In another family, a novel genomic fragment present in one child and absent in all other family members was observed. With the exception of this novel fragment, all polymorphic class I fragments observed in this study segregated with a serologically defined parental haplotype, a result consistent with HLA linkage.


Should providers of treatment be regarded as a random factor? If it ain't broke, don't "fix" it: a comment on Siemer and Joormann (2003).
In their criticism of B. E. Wampold and R. C. Serlin's analysis of treatment effects in nested designs, M. Siemer and J. Joormann argued that providers of services should be considered a fixed factor because typically providers are neither randomly selected from a population of providers nor randomly assigned to treatments, and statistical power to detect treatment effects is greater in the fixed than in the mixed model. The authors of the present article argue that if providers are considered fixed, conclusions about the treatment must be conditioned on the specific providers in the study, and they show that in this case generalizing beyond these providers incurs inflated Type I error rates.


Monitoring phosphatidic acid formation in intact phosphatidylcholine bilayers upon phospholipase D catalysis.
We have monitored the production of the negatively charged lipid, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidic acid acid (POPA), in supported lipid bilayers via the enzymatic hydrolysis of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (PC), a zwitterionic lipid. Experiments were performed with phospholipase D (PLD) in a Ca(2+) dependent fashion. The strategy for doing this involved using membrane-bound streptavidin as a biomarker for the charge on the membrane. The focusing position of streptavidin in electrophoretic-electroosmotic focusing (EEF) experiments was monitored via a fluorescent tag on this protein. The negative charge increased during these experiments due to the formation of POPA lipids. This caused the focusing position of streptavidin to migrate toward the negatively charged electrode. With the use of a calibration curve, the amount of POPA generated during this assay could be read out from the intact membrane, an objective that has been otherwise difficult to achieve because of the lack of unique chromophores on PA lipids. On the basis of these results, other enzymatic reactions involving the change in membrane charge could also be monitored in a similar way. This would include phosphorylation, dephosphorylation, lipid biosynthesis, and additional phospholipase reactions.


Contribution of area 17 to cell responses in the striate-recipient zone of the cat's lateral posterior-pulvinar complex.
The cat's lateral posterior-pulvinar complex (LP-pulvinar) contains three main representations of the visual field. The lateral part of the LP nucleus (LPl or striate-recipient zone) is the only region of these extrageniculate nuclei which receives afferents from the primary visual cortex. We investigated the contribution of area 17 to the response properties (orientation and spatial frequency tuning functions) of LPl neurons by cooling or lesioning the visual cortex. Responses of 40 LPl cells were studied before, during and after the reversible cooling of the striate cortex. When tested for orientation, a total of 10 units out of 28 was affected (36%). For most of these cells (eight of 10), cooling the visual cortex yielded a reduction of the cells' visual responses without altering their orientation-selectivity (there was no significant change in the orientation tuning width). For only two cells, inactivation led to an increase in the response amplitude. Also, blocking the visual cortex never modified the direction-selectivity of LPl cells. When tested for spatial frequency, 12 neurons out of 33 were affected (36%) by the experimental protocol. In most cases, we observed a reduction in the responses at each spatial frequency tested, with no change in tuning bandwidth. For only three LPl cells, the effects of inactivation of the visual cortex were restricted to specific spatial frequencies, altering the profile of the spatial frequency tuning function. In five cats, removing area 17 reduced the proportion of visual neurons in LPl and the spared visually evoked responses were noticeably depressed. Despite the reduction in responsiveness, a few LPl receptive fields within the cortical scotoma were still sensitive to the orientation and/or direction of a moving stimulus. This last observation suggests that some properties in LPl could be generated either by circuits intrinsic to the LPl or by afferents from extrastriate cortical areas. Overall, these results indicate that projections from the visual cortex to the striate-recipient zone of the LP-pulvinar complex are mainly excitatory. Despite the strong impact of the area 17 projections, our data suggest that the extrastriate cortex could also play a role in the establishment of response properties in the cat's LPl.


The "lazy-T" correction of ectropion of the lower punctum.
The "lazy-T" technique consists of a surgical horizontal and vertical shortening of the involved portion of the lower eyelid. As a result of the surgical procedure, the lid margin is restored to its normal relationship with the globe. The punctum is reestablished into its normal anatomical position in the proximity of the lacrimal lake. The cosmetic blemish resulting from the surgical scar is insignificant.


The selection of ELISA cut-off points for testing antibody to Newcastle disease by two-graph receiver operating characteristic (TG-ROC) analysis.
Two hundred and sixty field serum samples were tested for Newcastle disease (ND) antibodies using a commercial enzyme-linked immunosorbent assay (ELISA) and the haemagglutination inhibition test (HI). The HI test was regarded as the reference method. Reciprocal titres of 16 and above were considered positive. In this study the two-graph receiver operating characteristic (TG-ROC) analysis was used as a tool for selecting cut-off points. Sensitivity, specificity, efficiency and Youden's index were used as indices of test accuracy. The positive and negative predictive values of the ELISA results were analysed for various prevalence rates.


Retethering of sectioned fibrolipomatous filum terminales: report of two cases.
The release of a tethered spinal cord by sectioning a thickened filum terminale is a straightforward surgical procedure that can prevent, arrest, or ameliorate neurological deficits. We recently recognized progressive neurological deterioration caused by filum retethering in two patients years after this procedure was performed. This sequela of a recurrent tethered cord after the sectioning of a filum terminale has not previously been described. Two female patients, each 13 years of age at presentation, had been previously operated on for tethered spinal cords secondary to fibrolipomatous (fatty) fila terminale. Both presented with bladder dysfunction and one with progressive paraparesis. Magnetic resonance images revealed a low-lying conus medullaris and a sectioned filum with the proximal stump adherent to the posterior dura. Each patient underwent neurosurgical exploration of the previous site of sectioning, with the recognition of a retethered proximal stump of the filum terminale. After rerelease of the fatty filum, the patient with only bladder dysfunction stabilized and a motor examination revealed normal results for the patient with progressive paraparesis. Retethering of the spinal cord is a rare sequela after the sectioning of a tight filum terminale. The clinical presentation is typical for recurrent cord tethering, and the radiographic findings are subtle. Careful surgical exploration should be offered for spinal cord untethering. Awareness of this rare and hitherto undescribed sequela is necessary for appropriate long-term management of tethered spinal cord caused by a fatty filum terminale.


Selecting clinically relevant populations for reference intervals.
The selection of individuals to represent a group for any purpose can commit the effort to success or failure from the very outset. The description of this group must reflect what is, in reality, the purpose for which the group is being studied. Considering that reference intervals are designed to assist clinicians during diagnostic triage, criteria must reflect the nature of the subjects entering the system for care. The majority of people seeking care have non-life-threatening, yet troublesome complaints. In other words, persons who are in a reasonable state of health and who are not suffering from significant illness. Therefore the relevant cohort should fit this description, not individuals who lack any clinical or laboratory issues. Our focus is to employ reference intervals to exclude disease and not to define normality. Normality in the eyes of statisticians is quite different from the word as viewed by clinical staff, yet the term is used interchangeably without concern for its ultimate use. As a result medicine today stumbles over this issue at every turn. Scientists and artists do not speak the same language. Compounding this awkward situation has been the conviction that by increasing laboratory precision and accuracy, the utility of laboratory data, reference intervals in particular, will improve clinical understanding. This has, after many years, not proven to be the case. What has become very evident, however, is that unsustainably large sums of money are being devoted to improving laboratory performance. To what end, when we are unable to grasp the significance of what is available now including published reference intervals for common test values? A realistic perspective on the value of prevailing laboratory performance, as illustrated by reference intervals, must be adopted. The obligation of laboratories, its regulators and professional groups is to create a sustainable mechanism to provide clinically usable information, in a timely and affordable manner, to our population and to those legions of less fortunate humans.


Impact of Colonoscopy Preparation Video on Boston Bowel Preparation Scale Score.
Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the United States and the third most common cancer in men and the second most common cancer among women. Early detection of localized adenocarcinoma and adenomatous polyps helps reduce the mortality related to colon cancer. According to the American Gastroenterological Association, colonoscopy (CSPY) is the gold standard in screening for CRC. To improve the results of screening, the CSPY preparation (prep) needs to be optimal. This study was undertaken to determine whether a supplemental standardized educational video on bowel preparation in the viewer's native language would improve bowel preparation at the time of CSPY. After institutional review board approval, the records of adult patients who presented to the gastroenterology clinic were reviewed. Patients who underwent a CSPY were assigned according to whether they watched a supplemental educational video on CSPY bowel preparation in their native language. This video reflects the same information provided in written and verbal form at the time of CSPY scheduling. Bowel prep was rated by the endoscopist using the Boston Bowel Preparation Scale (BBPS) and quantifies the adequacy of the preprocedure bowel prep. Participant characteristics and BBPS scores were statistically assessed for significant differences. We identified a total of 186 patients, 91 in March 2015 (pre-video intervention) and 95 in March 2016 (post-video intervention). Mean BBPS score was 7.9 and 8.54 for the March 2015 and 2016 group, respectively (p value of .0039). Although there was no statistical difference between the 2 groups with concern to gender and age, the racial makeup and BBPS score were statistically different. Multivariate analysis was performed. There was no interaction between gender or race and year effect to account for any difference in that factors' performance. Thus, it can be implied that there is not a consistent race effect but there is a consistent gender effect with females having higher success rates, regardless of video intervention (p value of .003). After controlling for both gender and race, the year effect is modestly significant (p value of .025), with the post-video subjects having higher prep success rates. A supplemental educational video incorporated into precolonoscopy teaching may provide a standardized method of effectively conveying simple bowel prep instructions in an efficient manner. This study demonstrated that using such a video produced significant results in improving the quality of bowel preparation.


Fractional analysis of Th2-type cytokines in sequential samples of induced sputum.
Recent studies have demonstrated the usefulness of induced sputum in detecting the expression of Th2-type cytokines in asthmatics and have shown that the profile of inflammatory cells in induced sputum differs with time. To determine whether the duration of sputum induction also affects the expression of Th2-type cytokines in induced sputum. Induced sputum was collected from eight atopic asthmatics at two separated intervals (5 min each) during a 15 min sputum induction, and each sample was examined separately for cytokine expression and inflammatory cells. Using immunocytochemistry, interleukin (IL)-4 and IL-5 immunoreactivity, T lymphocytes (CD3), eosinophils (major basic protein), neutrophils (elastase), epithelial cells and macrophages (CD68) were compared in the induced sputum obtained from the 0 min to 5 min and the 10 min to 15 min samples. The number of immunoreactive-positive cells expressing IL-4 and IL-5 were significantly higher in the 10 to 15 min induced sputum sample than in the 0 min to 5 min induced sputum sample (P<0.05). The number of eosinophils was also significantly higher in the 10 min to 15 min sample than in the 0 min to 5 min sample (P<0.05). In contrast, the number of neutrophils was significantly higher in the 0 min to 5 min sample than in the 10 min to 15 min sample (P<0.01); T lymphocytes, macrophages and epithelial cells did not differ between the two samples. This study demonstrates that the duration of sputum induction significantly affects the profile of inflammatory cells and Th2-type cytokine expression, and underscores the need for the standardization of induced sputum procedure.


Direct access and control of the intracellular solution environment in single cells.
Methods that can control and vary the solution environment around single cells are abundant. In contrast, methods that offer direct access to the intracellular proteome and genome in single cells with the control, flexibility, and convenience given by microfluidic methods are both scarce and in great demand. Here, we present such a method based on using a microfluidic device mounted on a programmable scanning stage and cells on-chip permeabilized by the pore-forming glycoside digitonin. We characterized the on-chip digitonin poration, as well as the solution exchange within cells. Intracellular solution exchange times vary with the dose of exposure to digitonin from less than a second to tens of seconds. Also, the degree of permeabilization obtained for cells treated with the same dose varies considerably, especially for low doses of digitonin exposure and low permeabilities. With the use of the presented setup, the degree of permeabilization can be measured during the permeabilization process, which allows for "on-line" optimization of the digitonin exposure time. Using this calibrated permeabilization method, we demonstrate the generation of intracellular oscillations, intracellular gradients, and the delivery of substrate to initiate enzymatic reactions in situ. This method holds the potential to screen and titrate intracellular receptors or enzymes or to generate intracellular oscillations, useful in the study of signaling pathways and oscillation decoding among other applications.


Rho kinase polymorphism influences blood pressure and systemic vascular resistance in human twins: role of heredity.
The Rho/Rho kinase (ROCK) pathway is implicated in experimental hypertension. We, therefore, explored the role of ROCK2 genetic variation in human blood pressure (BP) regulation, exploiting the advantages of a human twin sample to probe heritability. The focus of this work is the common nonsynonymous variant at ROCK2: Thr431Asn. Cardiovascular and autonomic traits displayed substantial heritability (from approximately 33% to 71%; P<0.05). The Asn/Asn genotype (compared with Asn/Thr or Thr/Thr) was associated with greater resting systolic (P<0.001), diastolic (P<0.0001), and mean BP (P<0.0001); allelic variation at ROCK2 accounted for up to approximately 5% of BP variation (P<0.0001). Systemic vascular resistance was higher in Asn/Asn individuals (P=0.049), whereas cardiac output, large artery compliance, and vasoactive hormone secretion were not different. Coupling of the renin-angiotensin system to systemic resistance and BP was diminished in Asn/Asn homozygotes, suggesting genetic pleiotropy of Thr431Asn, confirmed by bivariate genetic analyses. The Asn/Asn genotype also predicted higher BP after environmental (cold) stress. The rise in heart rate after cold was less pronounced in Asn/Asn individuals, consistent with intact baroreceptor function, and baroreceptor slope was not influenced by genotype. Common genetic variation (Thr431Asn) at ROCK2 predicts increased BP, systemic vascular resistance (although not large artery compliance), and resistance in response to the endogenous renin-angiotensin system, indicating a resistance vessel-based effect on elevated BP. The results suggest that common variation in ROCK2 exerts systemic resistance-mediated changes in BP, documenting a novel mechanism for human circulatory control, and suggesting new possibilities for diagnostic profiling and treatment of subjects at risk of developing hypertension.


Regulation of prorenin secretion in cultured human transfected juxtaglomerular cells.
The regulation of renin secretion was studied in continuous culture of human juxtaglomerular cells (JGC), which provided a permanent source of human renin production (Pinet, F., M. T. Corvol, F. Dench, J. Bourguignon, J. Feunteun, J. Ménard, and P. Corvol, 1985, Proc. Natl. Acad. Sci. USA, 82:8503-8507). 95% of the renin species secreted was prorenin, and therefore this study concerned primarily prorenin secretion. Renin production was stable, since the cells had been maintained in culture for more than two years. In culture, these human cells formed colonies of smooth musclelike cells, and electron microscopy showed the presence of cytoskeleton structures including myofibrils and attachment bodies. This human model was used to investigate the control of prorenin secretion in vitro at cellular level. Various pharmacological agents known to stimulate or inhibit renin secretion were tested in the cell cultures. The variations in prorenin secretion were measured in the supernatant. Forskolin, an independent receptor activator of adenylate cyclase, stimulated prorenin secretion in a dose-dependent manner and this stimulation was mediated by 3',5' cyclic-AMP (cAMP). Angiotensin II (AII) was found to inhibit prorenin secretion directly in a dose-dependent manner and atrial natriuretic factor (ANF), whose effects on human JGC were characterized for the first time, was also shown to exert such inhibition. When the effects of this inhibition by AII and ANF were tested on forskolin-mediated stimulation of prorenin secretion, the latter was inhibited and no change occurred in cAMP release. When JGC were treated with histamine, bradykinin, or one or two bradykinin analogues, the responses suggested that in these cells, H2-histamine receptors and kinin receptors are dependent on adenylate cyclase. One peptide, substance P, had an inhibitory effect on prorenin secretion but it was less important than AII and ANF. The present results demonstrate that the adenylate cyclase system of human JGC remains intact during culture and supports the hypothesis that cAMP is the second messenger and Cai2+, the final messenger involved in renin secretion. The cell system used here permits the evaluation of cellular responses and intracellular events in granulated cells in a human model.


Undergraduate medical students' views about a reflective portfolio assessment of their communication skills learning.
To date, no studies have examined preclinical medical students' views about portfolios. Since portfolios are becoming increasingly valued in medical education, this study explores second-year medical students' views about a reflective portfolio assessment of their communication skills. 178 second-year medical students at the University of Nottingham completed the 18-item reflective portfolio questionnaire (RPQ) (alpha = 0.716) and a personal details questionnaire three days before submitting their portfolio assessment for communication skills. Data were analysed using univariate and multivariate statistics on SPSS Version 10.0. Total scores on the RPQ ranged from 40 to 75 (mean 58.28, SD 7.08). Significant relationships existed between RPQ total scores and students' ratings of their reflection skills (rs = 0.322, P < 0.001), RPQ total scores and students' confidence building another portfolio (T = 4.381, d.f. = 176, P < 0.001), and RPQ total scores and students' marks for their reflective portfolio assessment (rs = 0.167, P = 0.029). Students with more positive views about reflective portfolios were more likely to rate their reflection skills as good, receive better marks for their portfolio assessment, and be more confident building another portfolio. This study begins to highlight preclinical medical students' views about reflective portfolios. However, further research is required using qualitative studies to explore students' views in depth. Medical educators should be encouraged to consider introducing portfolios as a method of formative and summative assessment earlier in the medical curriculum.


A socio-technical systems approach to studying interruptions: understanding the interrupter's perspective.
The purpose of this study was to understand the cognitive processes underlying nurses' decision to interrupt other nurses. The Institute of Medicine (2000) reported that interruptions are likely contributors to medical errors. Unfortunately, the research to date has been quite homogenous, focusing only on the healthcare provider being interrupted, ignoring the true complexities of interruptions. This study took a socio-technical approach being the first to examine interruptions from the viewpoint of the interrupting nurse. Over 15 h of observations and 10 open-ended interviews with expert nurses in a Neuroscience Surgical Intensive Care Unit were conducted. It was found that nurses conduct a quick cost-benefit assessment to determine the interruptibility of other nurses and whether an interruption is value-added vs. non-value added. To complete the assessment, nurses consider several conditional factors related to the interruptee, the interrupter, and the nature of the interruption content, and different potential consequences of the interruption.


Expression of Osterix in mechanical stress-induced osteogenic differentiation of periodontal ligament cells in vitro.
Osterix (Osx) is an osteoblast-specific transcription factor required for the differentiation of pre-osteoblasts into functional osteoblasts. This study sought to examine the changes of Osx expression in periodontal ligament cells (PDLC) subjected to mechanical force, and to investigate whether Osx is involved in the mechanical stress-induced differentiation of PDLC. Human PDLC were exposed to centrifugal force for 1-12 h. Real-time polymerase chain reaction (PCR), western blot, and immunofluorescence assays were used to examine the mRNA and protein expression of Osx and its subcellular localization. Furthermore, PDLC were transfected with the expression vector pcDNA3.1 flag-Osx and subjected to mechanical force for 6 h. The changes in alkaline phosphatase (ALP) activity and in the expression of core-binding factor alpha1 (Cbfa1), ALP, osteopontin, bone sialoprotein, osteocalcin, and collagen I were measured. After the application of mechanical force, Osx was upregulated in a time-dependent manner at both mRNA and protein levels, and Osx protein was translocated from the cytosol into the cell nuclei. Overexpression of Osx did not affect the expression of Cbfa1, but it significantly enhanced the ALP activity and the mRNA expression of all the aforementioned osteogenic marker genes, all of which increased further under mechanical stress. These results suggest that Osx might play an important role in the mechanical stress-induced osteogenic differentiation of PDLC and therefore be involved in alveolar bone remodeling during orthodontic therapy.


[Correction of the antioxidative system during ozone therapy in peritonitis].
Clinical studies revealed correlation of changes in anti-oxidative system and lipid peroxidation with peritonitis phases. The assessment of anti-oxidative system was used as an additional diagnostic and prognostic criterion for the disease. Data are presented in proof of efficiency of an ozone physiological solution included in combined treatment of acute peritonitis. Ozone therapy helped to normalize functioning of the anti-oxidative defensive system and thereby decrease the incidence of severe complications and fatal cases among patients with peritonitis.


Clinical research using extracellular vesicles: insights from the International Society for Extracellular Vesicles 2018 Annual Meeting.
The abstracts presented at the 2018 International Society for Extracellular Vesicles Annual Meeting offer unique insight into the newest discoveries related to the biology and applied use of extracellular vesicles (EVs). As an extension of a recent "Clinical-Wrap Up" discussion at the International Society for Extracellular Vesicles 2018 Annual Meeting, a systematic review of each abstract was performed to determine which abstracts could be considered clinical research. Once the clinical research abstracts were identified, systematic data extraction included: the major focus of each clinical research abstract; the countries in which the work was done; and the sample size, if provided in the abstract. Each abstract was reviewed by two independent authors, with a third author resolving discrepancies in cases of disagreement. 174 out of 656 (27%) unique abstracts were determined to be clinical research. Oncology was a principal research focus (51 of the 174 clinical research abstracts, 29%). Many other clinical research abstracts presented at the International Society for Extracellular Vesicles 2018 Annual Meeting focused on the use of human samples for development of methods for potential application in the clinic. Beyond oncology and methods development, a wide range of topics was represented, including cardiovascular disease, neurodegenerative disease, genetics, and many others. Current research involving EVs highlights the common, but false dichotomy of science into curiosity-driven basic science or application-driven clinical research, when in fact both quest for understanding and intent to apply the findings appeared to drive much of the work at the International Society for Extracellular Vesicles 2018 Annual Meeting. Using Pasteur's Quadrant as a framework, we discuss where the field of EV research is heading and how we may gain insight into the biological function of EVs in tandem with how they may benefit individual health.


Relationship of Aortic Wall Distensibility to Mitral and Aortic Valve Calcification: The Multi-Ethnic Study of Atherosclerosis.
Data are limited on whether valvular calcification is associated with aortic wall stiffness. We tested whether aortic valve calcification (AVC) and/or mitral valve calcification (MVC) is inversely associated with aortic distensibility (AD). Cross-sectional study conducted in a subset of the Multi-Ethnic Study of Atherosclerosis (MESA) included 3676 MESA participants aged 44 to 84 years with AD measured with magnetic resonance imaging and with AVC and MVC measured with noncontrast cardiac computed tomography scans. Both AVC and MVC were divided into 3 categories: zero, < median values (low), and ≥ median values (high) for patients with nonzero values. Overall, 88% (n = 3256) and 92% (n = 3365) of participants had zero AVC and MVC, while 6% (n = 211) and 4% (n = 156) had low, and 6% (n = 209) and 4% (n = 155) had high values of AVC and MVC, respectively. The AVC was independently associated with AD after adjusting for age, gender, and ethnicity ( P = .035). No association was noted between AVC groups and AD after adjustment for all covariates or MVC groups and AD in any model.


Management of depression in the elderly.
Primary care physicians have a vital role to play in identifying depression in their elderly patients. Diagnosis may be difficult, because symptoms are atypical and frequently include psychomotor agitation, somatic symptoms, and complaints of memory loss. Patients with medical illnesses, such as cancer, postmyocardial infarction, stroke, Parkinson's disease, and early Alzheimer's disease are particularly vulnerable to depression. Drugs that may cause depressive symptoms are digitalis at toxic levels, beta-blockers, centrally acting antihypertensives, immunosuppressants, and nonsteroidal anti-inflammatory agents. Cyclic antidepressants are the drugs of first choice. Selection depends on the patient's physical health and current medications and the side effect profile of the drug. Side effects are more pronounced in old age because of drug accumulation owing to slowed clearance. Troublesome side effects are anticholinergic effects, orthostatic hypotension, sedation, cardiotoxicity, and weight gain. The most useful antidepressants for geriatric patients are the secondary amines, desipramine and nortriptyline. The second-generation drug trazodone has the advantage of causing the least anticholinergic effects, but it is very sedating. Before treatment, the patient should have an electrocardiogram, liver function tests, tonometry, sitting and standing blood pressures, evaluation of urinary symptoms for outflow obstruction, review of current medications, and estimation of suicide risk. Cyclic antidepressants are contraindicated during recovery from myocardial infarction, in heart disease when there is severe impairment of myocardial performance, in seizure disorders, and in the presence of glaucoma or a large prostate. Drug interactions that may cause trouble can occur with epinephrine, MAO inhibitors, thyroid hormone, cimetidine, and centrally acting antihypertensives. Dosage should start low, increasing usually by 25 mg every 4 to 5 days until a therapeutic level is reached. Failure of a noradrenergic antidepressant after 4 to 5 weeks can be followed by a trial of a serotonergic drug. Drug serum level monitoring is useful for imipramine, desipramine, and nortriptyline. Monoamine oxidase inhibitors are effective in many elderly patients who are resistant to TCAs. Sympathomimetic drugs must be avoided with MAOIs. Elderly patients are at high risk of toxicity and drug interactions with lithium. Electroconvulsive therapy is useful for patients who do not respond to drug treatment, but medical complications, particularly cardiovascular, often occur in patients 75 or older. Many patients relapse after ECT. Psychotherapy together with pharmacotherapy may be the optimal treatment for elderly depressives. Older patients are more likely to become chronically depressed than younger patients. The risk of suicide in depressed elderly males is high, particularly in those with psychosocial problems, and depression rises with age.


Impacts of gut bacteria on human health and diseases.
Gut bacteria are an important component of the microbiota ecosystem in the human gut, which is colonized by 1014 microbes, ten times more than the human cells. Gut bacteria play an important role in human health, such as supplying essential nutrients, synthesizing vitamin K, aiding in the digestion of cellulose, and promoting angiogenesis and enteric nerve function. However, they can also be potentially harmful due to the change of their composition when the gut ecosystem undergoes abnormal changes in the light of the use of antibiotics, illness, stress, aging, bad dietary habits, and lifestyle. Dysbiosis of the gut bacteria communities can cause many chronic diseases, such as inflammatory bowel disease, obesity, cancer, and autism. This review summarizes and discusses the roles and potential mechanisms of gut bacteria in human health and diseases.


[Analysis of methodological structure of research proposals submitted to the Health Care Research Fund].
Currently the methodological features of the applied-research grant proposals submitted to the Research Agencies are not well known while they play an important role in the evaluation process. The objectives of this work were to describe the methodological features and appropriateness in the fulfillment of a sample of proposals submitted to the Fondo de Investigación Sanitaria (FIS). 216 proposals submitted to the FIS in 1993 and 1994. A specific revision protocol was designed to assess the objectives of the study according with criteria drown from the scientific literature. Most of the proposals correctly fulfill the items relative to the research problem description and justification. The operative component of the scientific method is scarcely fulfilled in the proposal hypothesis, objectives and methods. A 30% of the proposals do not show a typical research frame. The observational design and the basic field research are the most frequent findings in their items. The study variables, subjects, information management, plan of analysis and organization are incomplete or wrong filled in more than 50% of the proposals. The accepted proposals had an score of 1.93 in the degree of appropriate fulfillment and the non accepted 1.76, p = 0.001. The item with the strongest association to the acceptation was the appropriate definition of the variables, OR = 21.6 (CI 95%: 4.90-45.7). The methodological structure is not appropriate in more than 50% of the proposals submitted to the FIS. The appropriateness in the fulfillment of the methodological features increase the probability of their acceptance.


Overexpression of the rice carotenoid cleavage dioxygenase 1 gene in Golden Rice endosperm suggests apocarotenoids as substrates in planta.
Carotenoids are converted by carotenoid cleavage dioxygenases that catalyze oxidative cleavage reactions leading to apocarotenoids. However, apocarotenoids can also be further truncated by some members of this enzyme family. The plant carotenoid cleavage dioxygenase 1 (CCD1) subfamily is known to degrade both carotenoids and apocarotenoids in vitro, leading to different volatile compounds. In this study, we investigated the impact of the rice CCD1 (OsCCD1) on the pigmentation of Golden Rice 2 (GR2), a genetically modified rice variety accumulating carotenoids in the endosperm. For this purpose, the corresponding cDNA was introduced into the rice genome under the control of an endosperm-specific promoter in sense and anti-sense orientations. Despite high expression levels of OsCCD1 in sense plants, pigment analysis revealed carotenoid levels and patterns comparable to those of GR2, pleading against carotenoids as substrates in rice endosperm. In support, similar carotenoid contents were determined in anti-sense plants. To check whether OsCCD1 overexpressed in GR2 endosperm is active, in vitro assays were performed with apocarotenoid substrates. HPLC analysis confirmed the cleavage activity of introduced OsCCD1. Our data indicate that apocarotenoids rather than carotenoids are the substrates of OsCCD1 in planta.


Preservation of normal gastric emptying following gastric surgery by use of a muscle bridge.
Maintenance of descending antral intramural pathways may be important in normal functioning of the pylorus after pylorus-preserving gastrectomy. We examined the effect of a 1-cm bridge of muscle as a means of maintaining pyloric connection to antral intramural motor control pathways in 10 pigs. Antropyloroduodenal pressures and transpyloric flow were measured during gastric emptying of saline before and after either total or subtotal transection and reanastomosis of antrum. Complete antral transection shortened the time interval between antral and subsequent pyloric lumen occlusion, significantly reducing total gastric emptying and volume of transpyloric flow pulses. Subtotal transection maintained pre-transection timing and was associated with normal patterns of transpyloric flow and emptying. Our results indicate that a muscle bridge is capable of maintaining normal gastric emptying and the coordination of antral with pyloric contractions. We propose that antropyloric coordination is maintained by transmission of neural signals through the muscle bridge.


Cross-Sectional Study of the Psychological Well-Being of Healthcare Workers in a Large European University Hospital after the COVID-19 Initial Wave.
The SARS-CoV-2 pandemic greatly impacted healthcare workers (HCWs) dedicated to caring for COVID-19 patients. The study was conducted in a large European hospital to study the psychological distress of HCWs engaged in COVID-19 wards in the early phase of the pandemic. A questionnaire was sent to 1229 HCWs aimed at collecting the following information: 1) sociodemographic data; 2) depression, anxiety, and stress scales (DASS-21); 3) event impact scale (IES-R); 4) perceived stress scale (PSS); and 5) work interface analysis. The responses were collected through Google® forms and then statistically analyzed. Regardless of the outcome of the questionnaire, all subjects were offered psychological support voluntarily. Approximately two-thirds of the workers reported no symptoms according to the DASS-21 scales, while the IES-R and PSS scales showed 36% and 43%, respectively. There were no statistically significant differences in the levels of depression investigated through the different scales in the various occupational categories. Symptoms of anxiety, stress, and depression were more pronounced in women, while the highest stress levels were observed in the younger age groups. The highest scores were observed on the DAS-21 scales of anxiety and IES-R but not on the others. Only 51 workers, most of them with previous SARS-CoV-2 infection, sought clinical psychological counseling, and more than half received subsequent psychological support. Our results agree with most of the literature data that anxiety, depression, and stress are associated with gender (female), age (18-44 vs. over 55), and having cared for patients with COVID-19.


National Survey: How Do We Approach the Patient at Risk of Clinical Deterioration outside the ICU in the Spanish Context?
Anticipating and avoiding preventable intrahospital cardiac arrest and clinical deterioration are important priorities for international healthcare systems and institutions. One of the internationally followed strategies to improve this matter is the introduction of the Rapid Response Systems (RRS). Although there is vast evidence from the international community, the evidence reported in a Spanish context is scarce. A nationwide cross-sectional research consisting of a voluntary 31-question online survey was performed. The Spanish Society of Intensive, Critical and Coronary Care Medicine (SEMICYUC) supported the research. We received 62 fully completed surveys distributed within 13 of the 17 regions and two autonomous cities of Spain. Thirty-two of the participants had an established Rapid Response Team (RRT). Common frequency on measuring vital signs was at least once per shift but other frequencies were contemplated (48.4%), usually based on professional criteria (69.4%), as only 12 (19.4%) centers used Early Warning Scores (EWS) or automated alarms on abnormal parameters. In the sample, doctors, nurses (55%), and other healthcare professionals (39%) could activate the RRT via telephone, but only 11.3% of the sample enacted this at early signs of deterioration. The responders on the RRT are the Intensive Care Unit (ICU), doctors, and nurses, who are available 24/7 most of the time. Concerning the education and training of general ward staff and RRT members, this varies from basic to advanced and specific-specialized level, simulating a growing educational methodology among participants. A great number of participants have emergency resuscitation equipment (drugs, airway adjuncts, and defibrillators) in their general wards. In terms of quality improvement, only half of the sample registered RRT activity indicators. In terms of the use of communication and teamwork techniques, the most used is clinical debriefing in 29 centers. In terms of the concept of RRS, we found in our context that we are in the early stages of the establishment process, as it is not yet a generalized concept in most of our hospitals. The centers that have it are in still in the process of maturing the system and adapting themselves to our context.


Epidermal growth factor receptor stimulation activates the RNA binding protein CUG-BP1 and increases expression of C/EBPbeta-LIP in mammary epithelial cells.
The transcription factor CCAAT/enhancer binding protein beta (C/EBP beta) is a key regulator of growth and differentiation in many tissues. C/EBP beta is expressed as several distinct protein isoforms (LAP1, LAP2, and LIP) whose expression is regulated by alternative translational initiation at downstream AUG start sites. The dominant-negative LIP isoform is predominantly expressed during proliferative cellular responses and is associated with aggressive tumors. In this study, we investigated a mechanism by which the LIP isoform is translationally regulated in mammary epithelial cells. We have demonstrated that LIP expression is increased in response to activation of the epidermal growth factor receptor (EGFR) signaling pathway and that the increased expression of LIP is regulated in part by an RNA binding protein referred to as CUG repeat binding protein (CUG-BP1). Our data demonstrate that EGFR signaling results in the phosphorylation of CUG-BP1 and this leads to an increase in the binding of CUG-BP1 to C/EBP beta mRNA and elevated expression of the LIP isoform. Phosphorylation is necessary for the binding activity of CUG-BP1 and the consequent increase in LIP expression, as determined by binding assays and a cell free, transcription-coupled translation system. CUG-BP1 is thus a previously unidentified downstream target of EGFR signaling and represents a new translational regulator of LIP expression in human mammary epithelial cells.


In vivo cholesterol synthesis by the rat digestive tract. II. A study of turnover.
In the main organs of the digestive tract of rats fed a semi-purified diet containing 0.5% cholesterol, cholesterol activity was measured 70 min and 8, 24 and 48 h after subcutaneous impulsion of 14C-acetate or intravenous injection of tritiated water. Cholesterol synthesized in the stomach and caecum-colon was not significantly renewed during the 48-hour experiment. On the contrary, cholesterol synthesized in situ in the intestine disappeared with a mean rate constant of 4% X h-1. The rate constant (K) varied (6% X h-1 in the duodenum and jejunum and about 3% X h-1 in the distal ileum) according to the site of the enterocytes in the small intestine. Cell sloughing could not account for the major part of the decrease in cholesterol radioactivity, particularly in the first three quarters of the small intestine. In the proximal half of the gut internal cholesterol secretion via lipoproteins poured into the lymph might play a role.


Electrospun amplified fiber optics.
All-optical signal processing is the focus of much research aiming to obtain effective alternatives to existing data transmission platforms. Amplification of light in fiber optics, such as in Erbium-doped fiber amplifiers, is especially important for efficient signal transmission. However, the complex fabrication methods involving high-temperature processes performed in a highly pure environment slow the fabrication process and make amplified components expensive with respect to an ideal, high-throughput, room temperature production. Here, we report on near-infrared polymer fiber amplifiers working over a band of ∼20 nm. The fibers are cheap, spun with a process entirely carried out at room temperature, and shown to have amplified spontaneous emission with good gain coefficients and low levels of optical losses (a few cm(-1)). The amplification process is favored by high fiber quality and low self-absorption. The found performance metrics appear to be suitable for short-distance operations, and the large variety of commercially available doping dyes might allow for effective multiwavelength operations by electrospun amplified fiber optics.


Endopeptidase-24.11 in pig lymph nodes. Purification and immunocytochemical localization in reticular cells.
Endopeptidase-24.11 (EC 22.214.171.124), a widely distributed cell-surface endopeptidase in pig tissues, was purified by immunoaffinity chromatography from its second most abundant source, lymph nodes. The detergent-solubilized enzyme is a glycoprotein with an apparent subunit Mr of 91,000, by electrophoresis in the presence of SDS. This value is intermediate between those observed in preparations from kidney and intestine. The specific activity (125I-labelled insulin B-chain as substrate) was similar to that prepared from other sources. Immuno-peroxidase and immunofluorescent cytochemical methods with either a monoclonal antibody, GK7C2, or an affinity-purified polyclonal antiserum, RP109, were used to establish the distribution and localization of the antigen in lymph nodes. Examination of many nodes confirmed the variability of endopeptidase-24.11 content from node to node. Pig lymph nodes are composed of functionally discrete nodelets and are anatomically inverted, with medulla being located peripheral to the cortex. Endopeptidase-24.11 was present in medulla, paracortex and cortex. The medulla, containing relatively few lymphocytes, stained more intensely than other zones. Lymphocyte-rich areas stained only weakly, but antigen was detectable in the centres of follicles and more strongly in a band surrounding them. The pattern of staining was reticular in appearance in all zones. In primary cell cultures, set up after enzymic disruption of nodes, the immuno-positive cells were found to be adherent to glass or plastic and to exhibit a fibroblastic morphology. Diffuse surface immunofluorescence and brighter intracellular immunofluorescence in granules were observed in these cells in the first few days of culture, but by the fourth day no immuno-positive cells remained and the fibroblasts that grew to confluence were somewhat different in morphology. The cells expressing the endopeptidase-24.11 antigen did not express Ia antigen and were clearly distinct from antigen-presenting dendritic cells. In appearance and properties they belong to the group described as reticular cells.


Are omega-3 fatty acids the most important nutritional modulators of coronary heart disease risk?
With each passing year, the evidence linking an increased risk for coronary heart disease (CHD) death with a chronic dietary deficiency in long-chain omega-3 (n-3) fatty acids (FAs) grows stronger. Recently, a federally mandated evidence-based review in the United States concluded that n-3 FAs, especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), have clear cardioprotective effects, and national and international expert panels and health organizations have begun to call for increased EPA and DHA intakes. Consumption of between 450 and 1000 mg/d is recommended for those without and with known CHD, respectively. Based on animal and isolated cell studies, these FAs were presumed to have antiarrhythmic effects. The first direct evidence for this in humans was recently published, as were new data linking low n-3 FA intakes with risk for developing atrial fibrillation. The strength of the n-3 story has now led to a proposal that blood levels of EPA plus DHA be considered a new, modifiable, and clinically relevant risk factor for death from CHD.


double-time is identical to discs overgrown, which is required for cell survival, proliferation and growth arrest in Drosophila imaginal discs.
We have isolated the discs overgrown gene of Drosophila and shown that it encodes a homolog of the Casein kinase I(delta)/(epsilon) subfamily and is identical to the double-time gene. However, in contrast to the weak double-time alleles, which appear to affect only the circadian rhythm, discs overgrown alleles, including bona fide null alleles, show strong effects on cell survival and growth control in imaginal discs. Analysis of their phenotypes and molecular lesions suggests that the Discs overgrown protein is a crucial component in the mechanism that links cell survival during proliferation to growth arrest in imaginal discs. This work provides the first analysis in a multicellular organism of Casein kinase I(delta)/(epsilon) functions necessary for survival. Since the amino acid sequences and three-dimensional structures of Casein kinase I(delta)/(epsilon) enzymes are highly conserved, the results suggest that these proteins may also function in controlling cell growth and survival in other organisms.


The cytokeratin expression of cultured human foetal keratinocytes.
Skin samples taken from foetuses of 15-19 weeks gestational age and keratinocytes were cultured by the 3T3 feeder method or in serum-free MCDB 153 medium on 16 mm coverslips. Keratinocytes taken from paediatric circumcisions and patients undergoing plastic surgery were also cultured using the 3T3 feeder method. A panel of monoclonal antibodies against a number of cytokeratins and differentiation markers were used in the PAP technique to analyse the cells. Cryostat sections taken from the donor skin samples were stained simultaneously. Foetal skin expressed the cytokeratins 7, 13 and 19 that were not observed postnatally. This cytokeratin expression as maintained in both culture conditions and also observe in paediatric and adult keratinocytes. Cytokeratin 7 was expressed on a greater proportion of foetal cells than in vivo, whereas the expression of 13 and 19 decreased. All keratinocytes expressed vimentin, transferrin receptor and Ki67 (proliferating cell antigen), while a small proportion expressed involucrin throughout culture, indicating their high level of differentiation and proliferation. No differences were observed between low and high density cultures. Foetal keratinocytes cultured in MCDB 153 did not reach confluence and stronger staining of differentiation markers was observed in these cells. These results show that the in vivo differences in cytokeratin expression of foetal and adult keratinocytes disappear in the culture.


Scandals in health-care: their impact on health policy and nursing.
Through an analysis of several high-profile scandals in health-care in the UK, this article discusses the nature of scandal and its impact on policy reform. The nursing profession is compared to social work and medicine, which have also undergone considerable examination and change as a result of scandals. The author draws on reports from public inquiries from 1945 to 2013 to form the basis of the discussion about policy responses following scandals in health-care. In each case, the nature of the scandal, the public and government discourses generated by events and the policy response to those failings are explored. These scandals are compared to the recent scandal at Mid Staffordshire Hospital. Conclusions are drawn about the impact of these events on the future of the profession and on health policy directions. Recent events have raised public anxieties about caring practices in nursing. Health policy reform driven by scandal may obscure the effect of under resourcing in health services and poses a very real threat to the continued support for state-run services. Understanding the socially constructed nature of scandal enables the nurse to develop a greater critical awareness of policy contexts in order that they can influence health service reform.


Palmitoylation is not required for trafficking of human anion exchanger 1 to the cell surface.
AE1 (anion exchanger 1) is a glycoprotein found in the plasma membrane of erythrocytes, where it mediates the electroneutral exchange of chloride and bicarbonate, a process important in CO2 removal from tissues. It had been previously shown that human AE1 purified from erythrocytes is covalently modified at Cys-843 in the membrane domain with palmitic acid. In this study, the role of Cys-843 in human AE1 trafficking was investigated by expressing various AE1 and Cys-843Ala (C843A) mutant constructs in transiently transfected HEK-293 cells. The AE1 C843A mutant was expressed to a similar level to AE1. The rate of N-glycan conversion from high-mannose into complex form in a glycosylation mutant (N555) of AE1 C843A, and thus the rate of trafficking from the endoplasmic reticulum to the Golgi, were comparable with that of AE1 (N555). Like AE1, AE1 C843A could be biotinylated at the cell surface, indicating that a cysteine residue at position 843 is not required for cell-surface expression of the protein. The turnover rate of AE1 C843A was not significantly different from AE1. While other proteins could be palmitoylated, labelling of transiently transfected HEK-293 cells or COS7 cells with [3H]palmitic acid failed to produce any detectable AE1 palmitoylation. These results suggest that AE1 is not palmitoylated in HEK-293 or COS7 cells and can traffic to the plasma membrane.


MTX intolerance in children and adolescents with juvenile idiopathic arthritis.
MTX is the medication most commonly used for antirheumatic treatment in juvenile idiopathic arthritis. It has high efficacy, is usually well tolerated and has an excellent safety profile. However, frequently intolerance symptoms develop that manifest as nausea, feelings of disgust or abdominal complaints prior to or directly after administration of the medication. No obvious toxicity is causing these intolerance symptoms, but symptoms are strictly limited to MTX and not transferred to other medications. MTX intolerance causes a significant reduction of quality of life in affected patients, frequently puts the treating physician in difficult situations regarding treatment choice, and may lead to uncomfortable decisions whether or not to stop an otherwise effective drug. Conventional countermeasures such as antiemetics, change of route from subcutaneous to oral or vice versa, or taste masking usually have only a limited effect. In this review, we present the current knowledge on MTX intolerance, its clinical picture and commonly employed strategies. We also consider newer behavioural treatment strategies that may offer a more effective symptom control.


Mode-locked 2 mum laser with highly thulium-doped silicate fiber.
We report self-starting passively mode-locked fiber lasers with a saturable absorber mirror using a piece of 30-cm-long newly developed highly thulium (Tm)-doped silicate glass fibers. The mode-locked pulses operate at 1980 nm with duration of 1.5 ps and energy of 0.76 nJ. This newly developed Tm-doped silicate fiber exhibits a slope efficiency of 68.3%, an amplified spontaneous emission spectrum bandwidth (FWHM) of 92 nm, and a gain per unit length of greater than 2 dB/cm. To the best of our knowledge, it is the first demonstration of mode-locked 2 mum fiber laser using shorter than 1-m-long active fiber, which paves the way for the demonstration of mode-locked fiber laser at 2 mum with gigahertz fundamental repetition rate.


Increased interaction of connexin43 with zonula occludens-1 during inhibition of gap junctions by G protein-coupled receptor agonists.
Astrocytes are extensively coupled through gap junctions (GJs) that are composed of channels mostly constituted by connexin43 (Cx43). This astroglial gap junctional intercellular communication (GJIC) allows propagation of ions and signaling molecules critical for neuronal activity and survival. It is drastically inhibited by a short-term exposure to endothelin-1 (ET-1) or to sphingosine-1-phosphate (S1P), both compounds being inflammatory mediators acting through activation of GTP-binding protein-coupled receptors (GPCRs). Previously, we have identified the GTPases G(i/o) and Rho as key actors in the process of S1P-induced inhibition. Here, we asked whether similar mechanisms underlied the effects of ET-1 and S1P by investigating changes in the phosphorylation status of Cx43 and in the molecular associations of Cx43 with zonula occludens (ZO) proteins and occludin. We showed that the inhibitory effect of ET-1 on GJIC was entirely dependent on the activation of G(i/o) but not on Rho and Rho-associated kinase. Both ET-1 and S1P induced dephosphorylation of Cx43 located at GJs through a process mediated by G(i/o) and calcineurin. Thanks to co-immunoprecipitation approaches, we found that a population of Cx43 (likely junctional Cx43) was associated to ZO-1-ZO-2-occludin multiprotein complexes and that acute treatments of astrocytes with ET-1 or S1P induced a G(i/o)-dependent increase in the amount of Cx43 linked to these complexes. As a whole, this study identifies a new mechanism of GJIC regulation in which two GPCR agonists dynamically alter interactions of Cx43 with its molecular partners.


[Malherbe's calcifying epithelioma--comment on two clinical cases].
Malherbe's calcifying epithelioma (also called trichomatricoma, pilomatrixoma or pilomatricoma) is an uncommon benign cutaneous tumor, which takes the form of a solitary, firm, asymptomatic, slowly growing nodule, developed subcutaneously on the face, neck, or proximal upper extremity. With an incidence of 1/800-1000 cutaneous tumors, pilomatricoma is more common in the first two decades of life, predominantly affecting the women. Multiple locations and associations between this entity and other diseases such as Steinert's myotonic dystrophy or Gardner syndrome have been described in the literature. The clinical diagnosis is often difficult, most cases being mistaken as sebaceous cysts. The treatment of choice is surgical excision. Incomplete surgical excision can be followed by local recurrence because most lesions are poorly defined; therefore, patients should be monitored after surgical excision to prevent this from happening. We present two cases admitted in 2010 and 2011 to the Iasi Department of Oral and Maxillo-Facial Surgery: a 55-year-old female patient with a Malherbe's calcifying epithelioma located in the left temporal region, and a 51-year-old female patient with the same disease but located in the left masseter region.


Sexual risk behavior among HIV-positive persons in Jamaica.
HIV/AIDS remains a global public health challenge, especially in sub-Saharan Africa and the Caribbean. Sexual risk behaviors among HIV-positive persons place their partners at risk for HIV transmission and other sexually transmitted infections. Stopping transmission acts among HIV-positive people is crucial in reversing HIV incidence. This study aimed to assess the prevalence and predictors of sexual risk behaviors among HIV-positive individuals in clinical care in Northwestern Jamaica. A cross-sectional survey of 118 (33 males and 85 females) HIV-positive individuals was used to assess demographic and health characteristics, HIV/AIDS knowledge, attitudes, and beliefs and sexual risk behaviors. About 12% of the study population stated that they had unprotected anal or vaginal sex without disclosing their HIV status. Participants who agreed that condoms reduce the risk of HIV transmission were 13.1 times more likely to use condoms during their last sexual encounters(95% CI: 2.1-79.0) than those who disagreed. About 75% of participants reported using a condom every time they had sexual intercourse in the past year, while 25% used condoms irregularly. Participants who had unprotected anal or vaginal sex without disclosing their status were less likely to have used condoms during the last sexual encounter (OR=0.1; 95% CI: 0.02-0.5). The prevalence of unsafe sex remains high among sexually active people living with HIV/AIDS in Jamaica. Study participants who engaged in unprotected sex without disclosing their HIV-positive status potentially place their partners at risk for HIV transmission and other sexually transmitted infections. The study findings highlight the need to promote safe sexual behaviors and a positive social environment for people living with HIV/AIDS in Jamaica.


Effects of the preoperative use of artificial tears combined with recombinant bovine basic fibroblast growth factor on cataract patients complicated with dry eyes.
To assess the effects of the preoperative application of artificial tears combined with recombinant bovine basic fibroblast growth factor on the ocular surface function and inflammatory factor levels after operation in cataract patients complicated with dry eyes. A total of 118 cataract patients (118 eyes) complicated with dry eyes treated from February 2019 to February2020 were assigned to control and observation groups (n=59 eyes/group) using a random number table. One week before the operation, the control group was administered 0.1% sodium hyaluronate eye drops (artificial tears), based on which the observation group received Beifushu eye drops (recombinant bovine basic fibroblast growth factor), both 6 times daily for 1 week. A comparison was made between the scores of clinical symptoms and the indices of ocular surface function, inflammatory factors in tears, and oxidative stress indices before and after the operation. The ocular surface function was evaluated by an ocular surface disease index questionnaire, tear film breakup-time assay, Schirmer's I test, and corneal fluorescein stain test. The inflammatory factors in tears were measured. No significant differences were noted in the general data and clinical symptom score, ocular surface disease index, tear film breakup-time, Schirmer's I test score, fluorescein stain score, interleukin-6, tumor necrosis factor-alpha, malondialdehyde, superoxide dismutase, lipid peroxide, and total antioxidant capacity before treatment between the 2 groups (p>0.05). After treatment, the clinical symptom score, ocular surface disease index, fluorescein stain score, tumor necrosis factor-alpha, interleukin-6, malondial-dehyde and lipid peroxide declined significantly, and tear film breakup-time, Schirmer's I test score, superoxide dismutase, and total antioxidant capacity increased in both the groups. The improvements in the clinical symptom score as well as in the indices of ocular surface function, inflammatory factors, and oxidative stress were more prominent in the observation group than in the control group (p<0.05). Artificial tears combined with recombinant bovine basic fibroblast growth factor before operation. significantly improved the ocular surface function, reduced inflammatory factors in tears, and alleviated dry eye symptoms after operation in cataract patients.


[The variability in the care for diabetic and hypertensive patients as a function of the styles of a physician's practice].
To analyse the variability of certain indicators of the procedure and outcome of medical care in the clinical records of diabetic and hypertense patients, and their relationship with the doctor's style of practice. An observational, descriptive study. Seven health centres in the public network. 40 primary care doctors (all the doctors). By means of stratified systematic random sampling, 20 clinical histories for each doctor were chosen (10 for patients diagnosed with diabetes mellitus and 10 with hypertension). The deviation ratio was used as the measurement of variability. Doctors' styles of medical practice were evaluated through the Australia questionnaire. Great variability was found in the frequency with which the different indicators of the two pathologies were recorded (deviation ratio: weight > 90.02% in DM, 81.37% in HT; height > 84.01% in DM, 77.10% in HT), and in the absolute figures (patients with HT, wide variability in all outcome indicators; patients with DM, in last figures, total and HDL cholesterol, and HbA1c). The styles of practice were shown to be related to the frequency of appearance of some indicators in the clinical records and with the quality of particular outcome indicators. The age of the doctor correlated significantly, and negatively, with the recording of almost all the indicators and their quality. Variability of medical practice in the care of the chronically ill seen in primary care does exist. The use of clinical practice guides is emphasised.


Mechanical stimulation by ultrasound enhances chondrogenic differentiation of mesenchymal stem cells in a fibrin-hyaluronic acid hydrogel.
Chondrogenic differentiation and cartilage tissue formation derived from stem cells are highly dependent on both biological and mechanical factors. This study investigated whether or not fibrin-hyaluronic acid (HA) coupled with low-intensity ultrasound (LIUS), a mechanical stimulation, produces an additive or synergistic effect on the chondrogenesis of rabbit mesenchymal stem cells (MSCs) derived from bone marrow. For the purpose of comparison, rabbit MSCs were first cultured in fibrin-HA or alginate hydrogels, and then subjected to chondrogenic differentiation in chondrogenic-defined medium for 4 weeks in the presence of either transforming growth factor-beta3 (TGF-β3) (10 ng/mL) or LIUS treatment (1.0 MHz and 200 mW/cm(2) ). The resulting samples were evaluated at 1 and 4 weeks by histological observation, chemical assays, and mechanical analysis. The fibrin-HA hydrogel was found to be more efficient than alginate in promoting chondrogenesis of the MSCs by producing a larger amount of sulfated glycosaminoglycans (GAGs) and collagen, and engineered constructs made with the hydrogel demonstrated higher mechanical strength. At 4 weeks of tissue culture, the chondrogenesis of the MSCs in fibrin-HA were shown to be further enhanced by treatment with LIUS, as observed by analyses for the amounts of GAGs and collagen, and mechanical strength testing. In contrast, TGF-β3, a well-known chondrogenic inducer, showed a marginal additive effect in the amount of collagen only. These results revealed that LIUS further enhanced chondrogenesis of the MSCs cultured in fibrin-HA, in vitro, and suggested that the combination of fibrin-HA and LIUS is a useful tool in constructing high-quality cartilage tissues from MSCs.


A frequency domain performance analysis of Horn and Schunck's optical flow algorithm for deformable motion.
A frequency domain performance analysis of Horn and Schunck's (1981) optical flow (HSOF) algorithm for estimation of deformable motion is presented. Noise sources in the algorithm are modeled using the discrete Fourier transform of the brightness pattern. This noise model along with the estimation error covariance function derived in previous work is used to derive an expression for the expected performance of the optical flow estimate that is valid for an arbitrary discrete brightness pattern. Simulation results are presented that demonstrate the validity of our methods and show that HSOF is more accurate that the optical flow estimate of Anandan for certain low-frequency patterns.


Eye movements evoked by microstimulation of dorsal cap of inferior olive in the rabbit.
1. Microstimulation was used in an attempt to activate selectively neurons of the dorsal cap of the inferior olive in unanesthetized rabbits, and the eye movements evoked by this microstimulation were recorded. 2. Trains of microstimulation (20--50 microA, 0.1- to 0.2-ms pulses, 10--60 pulses/s, 2--8 s duration) evoked low-velocity conjugate eye movements directed toward the side of the stimulated olive. These evoked eye movements were interrupted by resetting eye movements in the opposite direction and could be evoked only if the rabbit was in darkness. 3. The velocity of the evoked eye movements increased during a stimulus train and gradually decreased after the stimulation was terminated. 4. The low-velocity eye movements appeared to combine with eye movements evoked by vestibular stimulation without any significant interaction. 5. After transection of the olivocerebellar pathway or destruction of the contralateral cerebellar flocculus, dorsal cap microstimulation no longer evoked ipsilaterally directed eye movements.


Effect of vitamin D pretreatment of human mesenchymal stem cells on ectopic bone formation.
Adult mesenchymal stem cells (MSCs) are used in contemporary strategies for tissue engineering. The MSC is able to form bone following implantation as undifferentiated cells adherent to hydroxyapatite (HA)/tricalcium phosphate (TCP) scaffolds. Previous investigators have demonstrated that human MSCs (hMSCs) can be differentiated to osteoblasts in vitro by the inclusion of vitamin D and ascorbic acid. The aim of this study was to compare the osteogenic potential of predifferentiated and undifferentiated bone marrow-derived, culture-expanded hMSCs adherent to synthetic HA/TCP (60%/40%) following subcutaneous engraftment in severe combined immunodeficiency (SCID) mice. During the final 3 days of culture, cells were grown in Dulbecco's modified Eagle's medium containing 10% fetal calf serum and antibiotics or media containing 25-mM calcium supplementation with vitamin D and ascorbic acid. Four weeks following implantation in SCID mice, scoring analysis of bone formation within the cubes revealed the absence of bone formation in unloaded cubes. Bone formation compared by a qualitative bone index was 7.23% for undifferentiated cells compared to 5.20% for differentiated cells. Minimal resorption was observed at this early time point. In this ectopic model, predifferentiation using a combination of vitamin D and ascorbic acid failed to increase subsequent bone formation by implanted cells. Following implantation of hMSCs adherent to an osteoconductive scaffold, host factors may contribute dominant osteoinductive signals or impose inhibitory signals to control the fate of the implanted cell. Predifferentiation strategies require confirmation in vivo.


[Cardiovascular system in rats with varying resistance to acute hemorrhage].
Ultrasonic and laser doppler flowmetry, biomicroscopy were used in the study of systemic hemodynamics, microcirculation, blood flow in the brain, liver, kidneys and inferior limb in rats varying by resistance to acute blood loss. In the course of bleeding both high- and low-resistant rats (HRR, LRR) had the same decline in blood pressure, blood flow rate and tissue perfusion. When blood loss stopped. HRR developed relative compensation of the above parameters followed by irreversible fall. Posthemorrhagic recovery of these hemodynamic parameters in LRR was not observed. All the LRR showed persistent generalized constriction of hepatic, intestinal and renal microcirculation. Pumping performance of the heart continued to death in all HRR and in 65% of LRR. After stop of hemorrhage, 35% of LRR developed minor cardiac ejection syndrome which was followed by death within 30 min.


Comparing the Performance of Multiple Small-Data Personalized Tacrolimus Dosing Models for Pediatric Liver Transplant: A Retrospective Study.
Tacrolimus is a potent immunosuppressant used after pediatric liver transplant. However, tacrolimus's narrow therapeutic window, reliance on physicians' experience for the dose titration, and intra- and inter-patient variability result in liver transplant patients falling out of the target tacrolimus trough levels frequently. Existing personalized dosing models based on the area-under-the-concentration over time curves require a higher frequency of blood draws than the current standard of care and may not be practically feasible. We present a small-data artificial intelligence-derived platform, CURATE.AI, that uses data from individual patients obtained once daily to model the dose and response relationship and identify suitable doses dynamically. Retrospective optimization using 6 models of CURATE.AI and data from 16 patients demonstrated good predictive performance and identified a suitable model for further investigations.Clinical Relevance- This study established and compared the predictive performance of 6 personalized tacrolimus dosing models for pediatric liver transplant patients and identified a suitable model with consistently good predictive performance based on data from pediatric liver transplant patients.


Dithiolethiones inhibit NF-κB activity via covalent modification in human estrogen receptor-negative breast cancer.
The NF-κB transcription factor family influences breast cancer outcomes by regulating genes involved in tumor progression, angiogenesis, and metastasis. Dithiolethiones, a class of naturally occurring compounds with cancer chemoprevention effects that have become clinically available, have been found to inhibit NF-κB activity. However, the mechanism of this inhibition has not been identified, and the influence of dithiolethines on NF-κB pathway in breast cancer cells has not been examined. Here, we investigated the chemical and biochemical effects of dithiolethione on NF-κB and downstream effector molecules in estrogen receptor-negative breast cancer cells and murine tumor xenografts. The dithiolethiones ACS-1 and ACS-2 inhibited NF-κB transcriptional activity. Interestingly, this inhibition was not due to H(2)S release or protein phosphatase 2A activation, which are key properties of dithiolethiones, but occurred via a covalent reaction with the NF-κB p50 and p65 subunits to inhibit DNA binding. Dithiolethione-mediated inhibition of NF-κB-regulated genes resulted in the inhibition of interleukin (IL)-6, IL-8, urokinase-type plasminogen activator, and VEGF production. ACS-1 also inhibited matrix metalloproteinase-9 activity, cellular migration, and invasion, and ACS-2 reduced tumor burden and resulted in increased tumor host interactions. Together, our findings suggest that dithiolethiones show potential clinical use for estrogen negative breast cancer as a chemotherapeutic or adjuvant therapy.


Leukocyte filtration of the cardiotomy suction. Does it affect systemic leukocyte activation or pulmonary function?
Cardiopulmonary bypass is thought to propagate a global systemic response through contact with the non-physiological surfaces of the extracorporeal circuit, leading to the stimulation of leukocytes, their adherence to endothelial cells and the release of cytotoxic molecules. This, in turn, has been shown to accelerate pulmonary injury. This study tested a new leukocyte-filtration system (RemoweLL) against a conventional system with no leukocyte-depleting capacity to determine the efficacy of the filtration system and its effects on pulmonary function. Thirty patients underwent coronary artery bypass graft surgery using either the RemoweLL filtration system (15 patients) or a conventional cardiopulmonary bypass circuit (15 patients). Data were collected on the total number of leukocytes, their differentiation and activation, using the leukocyte adhesion integrin CD11b as a surrogate marker. Pulmonary function was assessed using the Alveolar-arterial Oxygenation Index (AaOI) and patients were categorized using the Berlin definition of acute respiratory distress syndrome (ARDS). Both groups showed significant increases in leukocyte numbers during CPB (p<0.001), with no differences noted between the groups. CD11b showed a significant increase in both groups, with peak activation occurring at the end of CPB, but no difference between the groups (p=0.8). There was a trend towards lower AaOI increases in the filtration group, but this did not reach significance (p=0.075) and there was no difference in ARDS definitions (p=0.33). Leukocyte filtration of cardiotomy suction did not influence total leukocyte counts or activation as measured by CD11b upregulation. Furthermore, no evidence could be found to suggest improved pulmonary function.