Self-renewal and multilineage differentiation of mouse dental epithelial stem cells.
Understanding the cellular and molecular mechanisms underlying the self-renewal and differentiation of dental epithelial stem cells (DESCs) that support the unlimited growth potential of mouse incisors is critical for developing novel tooth regenerative therapies and unraveling the pathogenesis of odontogenic tumors. However, analysis of DESC properties and regulation has been limited by the lack of an in vitro assay system and well-documented DESC markers. Here, we describe an in vitro sphere culture system to isolate the DESCs from postnatal mouse incisor cervical loops (CLs) where the DESCs are thought to reside. The dissociated cells from CLs were able to expand and form spheres for multiple generations in the culture system. Lineage tracing indicated that DESC within the spheres were epithelial in origin as evident by lineage tracing. Upon stimulation, the sphere cells differentiated into cytokeratin 14- and amelogenin-expressing and mineral material-producing cells. Compared to the CL tissue, sphere cells expressed high levels of expression of Sca-1, CD49f (also designated as integrin α6), and CD44. Fluorescence-activated cell sorting (FACS) analyses of mouse incisor CL cells further showed that the CD49f(Bright) population was enriched in sphere-forming cells. In addition, the CD49f(Bright) population includes both slow-cycling and Lgr5(+) DESCs. The in vitro sphere culture system and identification of CD49f(Bright) as a DESC marker provide a novel platform for enriching DESCs, interrogating how maintenance, cell fate determination, and differentiation of DESCs are regulated, and developing tooth regenerative therapies.


Conventional Ultrafiltration As Effective Strategy for Dye/Salt Fractionation in Textile Wastewater Treatment.
Use of tight ultrafiltration (UF) membranes has created a new pathway in fractionation of dye/salt mixtures from textile wastewater for sustainable resource recovery. Unexpectedly, a consistently high rejection for the dyes with smaller sizes related to the pore sizes of tight UF membranes is yielded. The potential mechanism involved in this puzzle remains unclear. In this study, seven tailored UF membranes with molecular weight cut-offs (MWCOs) from 6050 to 17530 Da were applied to separate dye/salt mixtures. These UF membranes allowed a complete transfer for NaCl and Na2SO4, due to large pore sizes. Additionally, these UF membranes had acceptably high rejections for direct and reactive dyes, due to the aggregation of dyes as clusters for enhanced sizes and low diffusivity. Specifically, the membrane with an MWCO of 7310 Da showed a complete rejection for reactive blue 2 and direct dyes. An integrated UF-diafiltration process was subsequently designed for fractionation of reactive blue 2/Na2SO4 mixture, achieving 99.84% desalination efficiency and 97.47% dye recovery. Furthermore, reactive blue 2 can be concentrated from 2.01 to 31.80 g·L-1. These results indicate that UF membranes even with porous structures are promising for effective fractionation of dyes and salts in sustainable textile wastewater treatment.


The genetics of dilated cardiomyopathy.
More than 40 different individual genes have been implicated in the inheritance of dilated cardiomyopathy. For a subset of these genes, mutations can lead to a spectrum of cardiomyopathy that extends to hypertrophic cardiomyopathy and left ventricular noncompaction. In nearly all cases, there is an increased risk of arrhythmias. With some genetic mutations, extracardiac manifestations are likely to be present. The precise genetic cause can usually not be discerned from the cardiac and/or extracardiac manifestations and requires molecular genetic diagnosis for prognostic determination and cardiac care. Newer technologies are influencing genetic testing, especially cardiomyopathy genetic testing, wherein an increased number of genes are now routinely being tested simultaneously. Although this approach to testing multiple genes is increasing the diagnostic yield, the analysis of multiple genes in one test is also resulting in a large amount of genetic information of unclear significance. Genetic testing is highly useful in the care of patients and families, as it guides diagnosis, influences care and aids in prognosis. However, the large amount of benign human genetic variation may complicate genetic results and often requires a skilled team to accurately interpret the findings.


Cigarette smoke extracts promote vascular smooth muscle cell proliferation and enhances contractile responses in the vasculature and airway.
Cigarette smoke exposure is a strong risk factor for cardiovascular and respiratory diseases. However, the knowledge about how cigarette smoke induces damage to vasculature and airway is limited. The present study was designed to examine the effects of cigarette smoke particles extracted by heptane (heptane-soluble smoke particles, HSP), by water (water-soluble smoke particles, WSP) and by DMSO (DMSO-soluble smoke particles, DSP), which represent lipophilic, hydrophilic and ambiphoteric constituents from the cigarette smoke, respectively. Human aortic smooth muscle cell (HASMC) proliferation was assessed in cell culture. Rat resistance artery and airway contractile responses to serotonin, U46619, phenylephrine, noradrenaline, acetylcholine, des-Arg⁹-bradykinin, bradykinin, sarafotoxin 6c and endothelin-1 were monitored by a sensitive myograph system. Immunocytochemistry and cell-based phosphoELISA assay were used to demonstrate activation of extracellular signal-regulated kinases 1/2 (ERK1/2). For the first time, our results demonstrate that although all the three extracts promote HASMC proliferation, the HSP and DSP effects occur earlier. HSP and DSP, but not WSP, increase the contractile responses to sarafotoxin 6c, U46619 or bradykinin in rat mesenteric artery and/or in bronchi. ERK1/2 is activated by HSP and DSP in HASMCs and inhibition of ERK1/2 abrogated the smoke extracts-induced HASMC proliferation, while blockage of nicotinic receptors had no effects, suggesting that the toxic effects of the smoke extracts occur via activation of intracellular ERK1/2 signalling, but not nicotinic receptors.


[Enhanced effectiveness of antibiotic therapy with an exogenous lysozyme in dysentery in children].
The course of dysentery and the host immune responsiveness were studied in 171 children with acute Flexner's and Sonne dysentery. 67 children were treated with monomycin in combination with lysozyme and 104 children with monomycin alone. It was shown that the recovery period in children treated with monomycin and lysozyme was shorter as compared to that in children treated with monomycin alone. The immunity characteristics in children treated with monomycin were closer to the physiological values. It is concluded that lysozyme potentiates the antibiotic therapy in children with dysentery. It should be used combined with monomycin in the treatment of children irrespective of the age and the disease pattern. The treatment course with the use of monomycin in combination with lysozyme is reduced to 5 days.


Differential protein profiling as a potential multi-marker approach for TSE diagnosis.
Transmissible spongiform encephalopathy describes a family of diseases affecting both man and animals. Current tests for the diagnosis of these diseases are based on the detection of an abnormal misfolded form of the host protein PrP which is found within the central nervous and lymphoreticular systems of affected animals. Recently, concern that this marker may not be as reliable as previously thought, coupled with an urgentneed for a pre-clinical live animal test, has led to the search for alternative assays for the detection of TSE disease. This "proof of concept" study, examines the use of differential protein expression profiling using surface enhanced laser desorption and ionisationtime of flight mass spectrometry (SELDI-TOF) for the diagnosis of TSE disease. Spectral output from all proteins selectively captured from individual murine brain homogenate samples, are compared as "profiles" in groups of infected and non-infected animals. Differential protein expression between groups is thus highlighted and statistically significant protein "peaks" used to construct a panel of disease specific markers.Studies at both terminal stages of disease and throughout the time course of disease have shown a disease specific protein profile or "disease fingerprint" which could be used to distinguish between groups of TSE infected and uninfected animals at an early time point of disease. Our results show many differentially expressed proteins in diseased and control animals, some at early stages of disease. Three proteins identified by SELDI-TOF analysis were verified by immunohistochemistry in brain tissue sections. We demonstrate that by combining the most statistically significant changes in expression, a panel of markers can be constructed that can distinguish between TSE diseased and normal animals. Differential protein expression profiling has the potential to be used for the detection of disease in TSE infected animals. Having established that a "training set" of potential markers can be constructed, more work would be required to further test the specificity and sensitivity of the assay in a "testing set". Based on these promising results, further studies are being performed using blood samples from infected sheep to assess the potential use of SELDI-TOF as a pre-mortem blood based diagnostic.


[The circumferential decompression by posterior transpedicular osteotomy and segmental instrumentation with interbody fusion for thoracic ossification of posterior iongitudinal ligament].
To observe the efficacy of the circumferential decompression with posterior transpedicular osteotomy and segmental instrumentation with interbody fusion for thoracic ossification of posterior Iongitudinal ligament (T-OPLL). From May 2012 to June 2015, 16 consecutive patients underwent posterior transpedicular osteotomy and segmental instrumentation with interbody fusion.Osteotomy range was depended by length and types of OPLL.Patient's data included level, clinical presentation, blood loss, length of surgery, complications, VAS, JOA, and Frankel grading system before and after the surgery. All data were collected, retrospectively. The follow-up period was (30±19) months (range from 12 to 50 months). The operation time was (261.6±51.3) min (range from 190 to 310 min). The blood loss was (980.3±370.5) ml (range from 600 to 2 100 ml). All patients were well treated with posterior compression and segmental instrumentation with interbody fusion.The VAS score was (4.2±0.2) in all patients at a week, improving to (2.7±0.1) points at 3 months, (2.4±0.2) at 1 year, and (2.0±0.1) at last fellow-up.The statistical analysis of the results showed a significant improvement of pain at 3 months (P<0.05) when compared to the preoperative status.The preoperative JOA score was (4.2±1.7) in all patients, improving to (7.8±2.5) points at 3 months, (8.5±2.7) at 1 year, and (9.0±1.0) at last fellow-up.The mean recovery rate for the total JOA score was (72%±8%). Differences in the overall JOA Scores showed significant postoperative improvement.Frankel grade improved by either 1 or 2 grades in 16 patients at the last follow-up.None of the patients showed any signs of instrument migration or failure during follow-up. The results suggested that the procedure achieved a total resection of the ossified posterior longitudinal ligament.The treatment method with posterior transpedicular osteotomy and circumferential decompression was found to be safe, effective, reliable, and technically feasible.


Drug Delivery Systems for Imaging and Therapy of Parkinson's Disease.
Although a variety of therapeutic approaches are available for the treatment of Parkinson's disease, challenges limit effective therapy. Among these challenges are delivery of drugs through the blood brain barier to the target brain tissue and the side effects observed during long term administration of antiparkinsonian drugs. The use of drug delivery systems such as liposomes, niosomes, micelles, nanoparticles, nanocapsules, gold nanoparticles, microspheres, microcapsules, nanobubbles, microbubbles and dendrimers is being investigated for diagnosis and therapy. This review focuses on formulation, development and advantages of nanosized drug delivery systems which can penetrate the central nervous system for the therapy and/or diagnosis of PD, and highlights future nanotechnological approaches. It is esential to deliver a sufficient amount of either therapeutic or radiocontrast agents to the brain in order to provide the best possible efficacy or imaging without undesired degradation of the agent. Current treatments focus on motor symptoms, but these treatments generally do not deal with modifying the course of Parkinson's disease. Beyond pharmacological therapy, the identification of abnormal proteins such as α -synuclein, parkin or leucine-rich repeat serine/threonine protein kinase 2 could represent promising alternative targets for molecular imaging and therapy of Parkinson's disease. Nanotechnology and nanosized drug delivery systems are being investigated intensely and could have potential effect for Parkinson's disease. The improvement of drug delivery systems could dramatically enhance the effectiveness of Parkinson's Disease therapy and reduce its side effects.


Weakly Supervised Adversarial Learning for 3D Human Pose Estimation from Point Clouds.
Point clouds-based 3D human pose estimation that aims to recover the 3D locations of human skeleton joints plays an important role in many AR/VR applications. The success of existing methods is generally built upon large scale data annotated with 3D human joints. However, it is a labor-intensive and error-prone process to annotate 3D human joints from input depth images or point clouds, due to the self-occlusion between body parts as well as the tedious annotation process on 3D point clouds. Meanwhile, it is easier to construct human pose datasets with 2D human joint annotations on depth images. To address this problem, we present a weakly supervised adversarial learning framework for 3D human pose estimation from point clouds. Compared to existing 3D human pose estimation methods from depth images or point clouds, we exploit both the weakly supervised data with only annotations of 2D human joints and fully supervised data with annotations of 3D human joints. In order to relieve the human pose ambiguity due to weak supervision, we adopt adversarial learning to ensure the recovered human pose is valid. Instead of using either 2D or 3D representations of depth images in previous methods, we exploit both point clouds and the input depth image. We adopt 2D CNN to extract 2D human joints from the input depth image, 2D human joints aid us in obtaining the initial 3D human joints and selecting effective sampling points that could reduce the computation cost of 3D human pose regression using point clouds network. The used point clouds network can narrow down the domain gap between the network input i.e. point clouds and 3D joints. Thanks to weakly supervised adversarial learning framework, our method can achieve accurate 3D human pose from point clouds. Experiments on the ITOP dataset and EVAL dataset demonstrate that our method can achieve state-of-the-art performance efficiently.


The interaction between the parkinsonian patient and his caregiver during feeding: a theoretical model.
A theoretical model for the assessment of the interaction between the patient with Parkinson's disease and his caregiver during assisted feeding is suggested. It is based on literature about impairments in Parkinson's disease and about interaction. The main concepts of the model are clarity of cues, sensitivity, interpretation, responsiveness and synchrony. It is suggested that the patient should be allowed to focus on the socio-emotional content versus the task content of communication during different phases of interaction during the meal.


A hyperfusogenic gibbon ape leukemia envelope glycoprotein: targeting of a cytotoxic gene by ligand display.
An important goal in cancer gene therapy is the development of novel targeted cytotoxic genes. The observation that transfection of a GaLV envelope glycoprotein lacking an R peptide into human cells results in considerable cell-cell fusion and subsequent cell death prompted us to explore the potential for using this fusogenic membrane glycoprotein (FMG) as a targeted cytotoxic gene. As proof of principle, we therefore displayed epidermal growth factor (EGF) on the N terminus of GaLV envelope glycoproteins both with and without an R peptide (GaLV R+ and GaLV R-). Transfection of the GaLVR+ envelope expression plasmids did not cause cell-cell fusion. The GaLV R+ envelopes were incorporated into retroviral vectors whose infectivity was investigated on EGF receptor-positive and -negative cells. The vector incorporating an N-terminally unmodified envelope was able to infect all human cell lines tested. Infectivity of the vector incorporating an envelope on which EGF was displayed was restricted on EGF receptor-positive cells (but not on EGF receptor-negative cells) and could be restored by protease cleavage of the displayed domain or competition with exogenous ligand. The cell-cell fusion capacity of the GaLV R- envelope glycoproteins (N-terminally unmodified and with N-terminal display of both EGF and insulin-like growth factor I [IGF-I]) was investigated by plasmid DNA transfection. While the N-terminally unmodified GaLV R- fused all human cell types tested, fusogenicity of GaLV R- on which EGF or IGF-I was displayed was considerably restricted on receptor-positive cells. "Reciprocal" competition experiments showed that fusogenicity could be restored by competition only with the relevant exogenous ligand. Thus the specificity of cell-cell fusion by a hyperfusogenic GaLV envelope glycoprotein can be regulated by N-terminal display of growth factor ligands. There is therefore significant potential for further development of the targeting of the cell-killing capability of this fusogenic viral glycoprotein by using strategies similar to those we have developed for the targeting of retroviral vectors.


Patterned Illumination Techniques in Optogenetics: An Insight Into Decelerating Murine Hearts.
Much has been reported about optogenetic based cardiac arrhythmia treatment and the corresponding characterization of photostimulation parameters, but still, our capacity to interact with the underlying spatiotemporal excitation patterns relies mainly on electrical and/or pharmacological approaches. However, these well-established treatments have always been an object of somehow heated discussions. Though being acutely life-saving, they often come with potential side-effects leading to a decreased functionality of the complex cardiac system. Recent optogenetic studies showed the feasibility of the usage of photostimulation as a defibrillation method with comparatively high success rates. Although, these studies mainly concentrated on the description as well as on the comparison of single photodefibrillation approaches, such as locally focused light application and global illumination, less effort was spent on the description of excitation patterns during actual photostimulation. In this study, the authors implemented a multi-site photodefibrillation technique in combination with Multi-Lead electrocardiograms (ECGs). The technical connection of real-time heart rhythm measurements and the arrhythmia counteracting light control provides a further step toward automated arrhythmia classification, which can lead to adaptive photodefibrillation methods. In order to show the power effectiveness of the new approach, transgenic murine hearts expressing channelrhodopsin-2 ex vivo were investigated using circumferential micro-LED and ECG arrays. Thus, combining the best of two methods by giving the possibility to illuminate either locally or globally with differing pulse parameters. The optical technique presented here addresses a number of challenges of technical cardiac optogenetics and is discussed in the context of arrhythmic development during photostimulation.


Renal tumors in young adults.
The clinical and pathological features of solid or complex cystic renal masses in young adults have not been defined. We present our experience with patients 17 to 45 years old with such renal masses to define the incidence of malignant vs benign lesions, familial tendencies and clinical outcomes. The medical records of all patients 17 to 45 years old who presented with a solid or suspicious complex cystic renal mass at 2 tertiary care hospitals between 1988 and 2002 were retrospectively reviewed. Pertinent clinical information was compiled, including age, gender, mode of presentation, renal function, year and type of surgery, pathological analysis and survival data. There were 114 evaluable patients who underwent a total of 119 nephrectomies. Mean patient age was 37.1 years and males comprised 56.1% of the population. Twelve patients had familial renal cell carcinoma (RCC), the von Hippel-Lindau syndrome. Mode of presentation for patients with sporadic disease was symptomatic (55.9%), incidental (35.3%) or unknown (8.8%). Radical nephrectomy, partial nephrectomy and nephroureterectomy were performed in 80 kidneys (67.2%), 37 (31.1%) and 2 (1.7%), respectively. Malignant lesions comprised 79.8% of all masses and 95.8% of these were renal cell carcinoma. Of the RCCs 75.8% were grade 1 or 2 and 89% were organ confined. Young women were much more likely than men to have a benign lesion (36.0% vs 9.5%, p <0.01) and the diversity of histologies was impressive (of the 24 total benign masses 9 were different tumor types). With an average followup of 38.3 months overall survival is 90.2%. Among patients with RCC 84.9% are alive and cancer-free, 11.6% are dead from disease and 3.5% are alive with recurrent disease. We report the largest known series of solid or suspicious complex renal masses in young adults. As expected, familial tumors are more common in this population. While RCC is the most common tumor, a wide variety of potential pathological outcomes are possible, particularly in women, who were much more likely to have a benign lesion. RCC in this patient population appears to have a favorable prognosis, despite symptomatic presentation in the majority of cases.


Review of strategies to recruit smokers for smoking cessation: a population impact perspective.
This paper reviews published articles describing several instruments used currently to "capture" the attention of smokers for quit-smoking interventions and emphasizes the distinction between the strategies used to reach eligible individuals and those used to recruit them for the proposed smoking cessation intervention. The search for articles was conducted using MEDLINE, PsychARTICLE, and LILACS. Key words for the search included recruitment, enrollment, reach, smoking cessation, quitline, and helpline mentioned in the abstracts and titles of the articles. Articles published in English, Portuguese, and Spanish through November 2006 that emphasized capture instruments related to different populations and specific interventions were included in this review. Twenty-nine studies met the inclusion criteria. Studies that used active and mixed strategies reached, on average, a smaller and less diverse possible number of eligible individuals and had greater participation proportions than those that used reactive strategies. Future studies are needed to evaluate the effectiveness of new associations between current interventions and reactive capture strategies, in view of the potential for increasing the population impact related to reactive capture.


Preadult stage parasites and multiple timed exposure to infective larvae are involved in development of limb edema in Brugia malayi-infected Indian leaf monkeys (Presbytis entellus).
The pathogenesis of filarial limb edema is not known. The role of parasitological variables and parasite-mediated phenomena in the development of limb edema was investigated in the Presbytis entellus-Brugia malayi model. Infection was initiated with subcutaneous inoculation of infective third-stage larvae (L(3)), and the animals were reexposed to different doses of L(3) at the prepatent, patent, and diminishing microfilaremia (0 to 5% of peak microfilaremia count) stages of infection. A large L(3) inoculum size and repeated inoculation in the ankle region during the prepatent, patent, and diminishing microfilaremia stages of infection were found to be necessary for reproducible induction of limb edema. The preadult stage of the parasite was found to be the most potent inducer of limb edema, followed by L(5) and L(4). The presence of the proinflammatory cytokines tumor necrosis factor alpha, interleukin-1beta, and interleukin-6 in edema fluid in the leg receiving the parasite challenge indicated that the limb edema development was due to parasite-mediated cytokine responses. The absence of bacterial infection or anti-streptolysin O titer in the edema fluid and blood indicated that bacterial infection is not necessary for the development of limb edema.


Effects of Inner Nuclear Membrane Proteins SUN1/UNC-84A and SUN2/UNC-84B on the Early Steps of HIV-1 Infection.
Human immunodeficiency virus type 1 (HIV-1) infection of dividing and nondividing cells involves regulatory interactions with the nuclear pore complex (NPC), followed by translocation to the nucleus and preferential integration into genomic areas in proximity to the inner nuclear membrane (INM). To identify host proteins that may contribute to these processes, we performed an overexpression screen of known membrane-associated NE proteins. We found that the integral transmembrane proteins SUN1/UNC84A and SUN2/UNC84B are potent or modest inhibitors of HIV-1 infection, respectively, and that suppression corresponds to defects in the accumulation of viral cDNA in the nucleus. While laboratory strains (HIV-1NL4.3 and HIV-1IIIB) are sensitive to SUN1-mediated inhibition, the transmitted founder viruses RHPA and ZM247 are largely resistant. Using chimeric viruses, we identified the HIV-1 capsid (CA) protein as a major determinant of sensitivity to SUN1, and in vitro-assembled capsid-nucleocapsid (CANC) nanotubes captured SUN1 and SUN2 from cell lysates. Finally, we generated SUN1-/- and SUN2-/- cells by using CRISPR/Cas9 and found that the loss of SUN1 had no effect on HIV-1 infectivity, whereas the loss of SUN2 had a modest suppressive effect. Taken together, these observations suggest that SUN1 and SUN2 may function redundantly to modulate postentry, nuclear-associated steps of HIV-1 infection.IMPORTANCE HIV-1 causes more than 1 million deaths per year. The life cycle of HIV-1 has been studied extensively, yet important steps that occur between viral capsid release into the cytoplasm and the expression of viral genes remain elusive. We propose here that the INM components SUN1 and SUN2, two members of the linker of nucleoskeleton and cytoskeleton (LINC) complex, may interact with incoming HIV-1 replication complexes and affect key steps of infection. While overexpression of these proteins reduces HIV-1 infection, disruption of the individual SUN2 and SUN1 genes leads to a mild reduction or no effect on infectivity, respectively. We speculate that SUN1/SUN2 may function redundantly in early HIV-1 infection steps and therefore influence HIV-1 replication and pathogenesis.


PEAKS DB: de novo sequencing assisted database search for sensitive and accurate peptide identification.
Many software tools have been developed for the automated identification of peptides from tandem mass spectra. The accuracy and sensitivity of the identification software via database search are critical for successful proteomics experiments. A new database search tool, PEAKS DB, has been developed by incorporating the de novo sequencing results into the database search. PEAKS DB achieves significantly improved accuracy and sensitivity over two other commonly used software packages. Additionally, a new result validation method, decoy fusion, has been introduced to solve the issue of overconfidence that exists in the conventional target decoy method for certain types of peptide identification software.


Discharge of respiratory-compromised children after respiratory rehabilitation.
There is a growing demand for respiratory rehabilitation services for children dependent on tracheostomy and/or chronic mechanical ventilation. Discharging these patients home following their rehabilitation can be an arduous process. To define the length of time required to rehabilitate and discharge these patients, and to identify predictors of a prolonged or failed discharge process. We conducted a retrospective chart review of patients admitted to the Respiratory Rehabilitation Unit at Alyn Hospital, Jerusalem, over a 4 year period. Of the 48 patients identified, 31 (64.7%) were eventually discharged, 13 (27.1%) remained hospitalized long-term, and 4 (8.3%) died during their hospitalization. The median length of hospitalization was 10 months: 6 months for purposes of rehabilitation therapy, and 4 months thereafter to resolve the logistics of discharge. Specific family characteristics--an unemployed father (odds ratio = 4.6, P = 0.02) and an additional family member with a disability (OR = 5.8, P = 0.03)--as well as ongoing mechanical ventilation at the time of discharge (OR = 5.5, P < 0.01) were found to positively correlate with a prolonged or failed discharge process. Hospitalization in a pediatric respiratory rehabilitation unit may be prolonged for both medical and non-medical reasons, with the process of discharge home being particularly difficult in certain subsets of patients. A proactive discharge policy by hospitals, improved community support services, and legislation defining the rights of home-ventilated children may facilitate more efficient discharge home of these patients.


Unusual relaxation pathway from the two-photon excited first singlet state of carotenoids.
Transient infrared and visible absorption measurements along with density functional theory (DFT) calculations on carotenoids 8'-apo-beta-caroten-8'-al (I) and 7',7'-dicyano-7'-apo-beta-carotene (II) were used to explore the nature of a long-lived species observed in transient infrared absorption measurements following two-photon excitation (Pang et al. J. Phys. Chem. B 2009, 113, 13806). The long-lived species of I has a very strong infrared absorption around 1510 cm(-1) and a visible transient absorption band centered at 760 nm. The long-lived species appears on two different time scales of approximately 16 and 140-270 ps. The longer rise component is absent in nonpolar solvents. DFT calculations using the B3LYP functional and the 6-31G(d) basis set were used to investigate the ground-state potential-energy surface of I and II including its conformational isomers, a pi-diradical "kinked" structure, and cation and neutral radicals. From the simulated infrared spectra of all the structures considered, we found a close match in the cation radical spectrum to the experimental infrared spectrum of the long-lived species. However, the visible absorption band does not match that of the monomeric cation radical. On the basis of our experimental and theoretical results, we propose a charge-transfer complex between a carotenoid and a solvent molecule for the origin of the long-lived species formed from the direct two-photon excitation of the S(1) state.


Alteration in the Cytokine Secretion of Bone Marrow Stromal Cells from Patients with Chronic Myelomonocytic Leukemia Contribute to Impaired Hematopoietic Supportive Activity.
Bone marrow stromal cells (BMSCs) represent an important cellular component of the bone marrow microenvironment, which play an important role in supporting and regulating the proliferation and differentiation of hematopoietic stem/progenitor cells (HSPCs). We have previously reported that the ability of BMSCs derived from CMML patients (CMML-BMSCs) in supporting the expansion of cord blood (CB) CD34+ cells was significantly reduced compared to BMSCs derived from healthy donors (HD-BMSCs). In addition, CMML-BMSCs led to a skewed differentiation of CB CD34+ cells favoring myeloid lineage compared with HD-BMSCs. To assess whether the altered cytokine secretion was one of the mechanisms to mediate the impaired hematopoietic supportive activity of CMML-BMSCs, a transwell coculture followed by cytokine array was performed. We showed that noncontacted coculture with CMML-BMSCs preferentially promoted the differentiation of CB CD34+ cells toward myeloid lineage. The expression levels of multiple cytokines (IL-6, IL-8, and GRO-β) were markedly reduced in CMML-BMSCs compared with HD-BMSCs. By supplementing IL-6, IL-8, or GRO-β, the hematopoietic supportive activity of CMML-BMSCs was partially restored. These results suggested that BMSCs may contribute to the pathogenesis of CMML by altering their cytokine secretion, which will shed light on the further investigation to develop novel therapeutic strategies for CMML patients.


Does elevated hemoglobin F modulate the phenotype in Hb SD-Los Angeles?
Hemoglobin (Hb) SD-Los Angeles compound heterozygotes usually have a severe clinical course although the effect of an elevated Hb F on the clinical phenotype has not been reported previously. We describe 5 Kuwaiti children with Hb SD associated with Hb F levels >20%. They all presented with sickling-related symptoms by < or =2 years of age and have been followed for 3-15 years. All had severe clinical courses marked by varying degrees of splenic sequestration crises, acute chest syndrome, vaso-occlusive crises, osteomyelitis and avascular necrosis of the femoral head. This pattern is in contrast with the usually mild presentation in Kuwaiti Hb SS patients with elevated Hb F. It therefore appears that Hb F does not ameliorate the clinical phenotype in Hb SD-Los Angeles. The reasons for this are not quite clear.


Nano-hydroxyapatite particles induce apoptosis on MC3T3-E1 cells and tissue cells in SD rats.
While the advantages of nanomaterials are being increasingly recognized, their potential toxicity is drawing more and more attention and concern. In this study, we explore the toxicity mechanism of 20-30 nm rod-shaped hydroxyapatite (HA) nanoparticles in vitro and in vivo. The nanoparticles were prepared by precipitation and characterized by IR, XRD and TEM. Concentrations of 0 μg mL(-1), 10 μg mL(-1), 100 μg mL(-1), 1 mg mL(-1), and 10 mg mL(-1) were applied to the MC3T3-E1 cells for viability (MTT-test). Based on the characteristic differences of the two methods of cell death, the morphological features of the MC3T3-E1 cell line co-cultured with nano-hydroxyapatite (n-HA) (10 mg mL(-1)) for 24 h were also observed by TEM. Furthermore, important serum biochemical markers and histopathological examinations were used to evaluate the potential toxicological effect of n-HA on the major organs of SD rats injected intraperitoneally with n-HA (33.3 mg kg(-1) body weight). In the results, we found cell growth inhibition and apoptosis in MC3T3-E1 cells co-cultured with n-HA. Moreover, apoptosis but not necrosis was illustrated in liver and renal tissue by using histopathology slices and serum biochemical markers. It suggests that apoptosis may be the possible mechanism of n-HA toxicity and provides a better understanding of the biocompatibility of nanomaterials applied in human bone repair.


Phenotypic characterization of a copA mutant of Neisseria gonorrhoeae identifies a link between copper and nitrosative stress.
NGO0579 is annotated copA in the Neisseria gonorrhoeae chromosome, suggesting that it encodes a cation-transporting ATPase specific for copper ions. Compared to wild-type cells, a copA mutant was more sensitive to killing by copper ions but not to other transition metals. The mutant also accumulated a greater amount of copper, consistent with the predicted role of CopA as a copper efflux pump. The copA mutant showed a reduced ability to invade and survive within human cervical epithelial cells, although its ability to form a biofilm on the surface of these cells was not significantly different from that of the wild type. In the presence of copper, the copA mutant exhibited increased sensitivity to killing by nitrite or nitric oxide. Therefore, we concluded that copper ion efflux catalyzed by CopA is linked to the nitrosative stress defense system of Neisseria gonorrhoeae. These observations suggest that copper may exert its effects as an antibacterial agent in the innate immune system via an interaction with reactive nitrogen species.


Treatment of Knee Osteochondritis Dissecans With a Cell-Free Biomimetic Osteochondral Scaffold: Clinical and Imaging Findings at Midterm Follow-up.
Osteochondritis dissecans (OCD) is a developmental condition of subchondral bone that may result in secondary separation and instability of the overlying articular cartilage, which in turn may lead to degeneration of the overall joint and early osteoarthritis. Biphasic scaffolds have been developed to address defects of the entire osteochondral unit by reproducing the different biological and functional requirements and guiding the growth of both bone and cartilage. To evaluate midterm clinical and imaging results after cell-free osteochondral scaffold implantation for the treatment of knee OCD. Case series; Level of evidence, 4. Twenty-seven patients (8 women, 19 men; mean age, 25.5 ± 7.7 years) were treated for knee OCD, with International Cartilage Repair Society (ICRS) grade 3 to 4 lesions with a mean size of 3.4 ± 2.2 cm2 (range, 1.5-12 cm2), and prospectively evaluated for up to 5 years using the ICRS classification system and the Tegner score. Eighteen patients underwent magnetic resonance imaging (MRI) at 24 and 60 months of follow-up, and the graft was evaluated using the magnetic resonance observation of cartilage repair tissue (MOCART) score for the cartilage layer, while a specific score was used for subchondral bone. All patients significantly improved their clinical scores at each follow-up until their final evaluation. The mean International Knee Documentation Committee (IKDC) subjective score improved from 48.4 ± 17.8 to 82.2 ± 12.2 at 2 years ( P < .0005), and it then remained stable for up to 5 years postoperatively (90.1 ± 12.0). The mean Tegner score increased from 2.4 ± 1.7 preoperatively to 4.4 ± 1.6 at 2 years ( P = .001), with a further increase up to 5.0 ± 1.7 at 5 years of follow-up ( P < .0005 vs preoperatively), reaching almost the preinjury level (5.7 ± 2.2). The MOCART score showed stable results between 24 and 60 months, whereas the subchondral bone status significantly improved over time. No correlation was found between MRI findings and clinical outcomes. This 1-step cell-free scaffold implantation procedure showed good and stable results for up to 60 months of follow-up for the treatment of knee OCD. MRI showed abnormalities, in particular at the subchondral bone level, but there was an overall improvement of features over time. No correlation was found between imaging and clinical findings.


Zinc determined in 10-microL serum or urine samples by flameless atomic absorption spectrometry.
We describe a precise flameless atomic absorption spectrometric method requiring only 10 microL of sample, which thus permits repeated measurements in the neonate. Standard curves covering the range of 0 to 30 mg of zinc per liter, with standards in various matrixes (albumin, glycerol, serum, or nitric acid) had slopes ranging from 0.49 to 1.17, relative to that for aqueous standards. We prepared low-zinc matrixes, which had slopes similar to that of serum, by dialyzing serum or a 50 g/L solution of albumin vs a buffered zinc-free dialysis fluid containing appropriate inorganic constituents. Use of thoroughly de-ionized water, reagent-grade chemicals, "ultrapure" nitric acid, specified disposable plastic ware, and appropriate pipet tip-rinsing techniques minimized extraneous contamination with zinc. Concentrations of zinc in serum calculated from a "rational-method" calibration algorithm fit to the standard curve agreed well with independent determinations by flame atomic absorption spectrometry.


Promoting South Asian primary care research and projecting family medicine practitioner-scholars: A matter of critical importance.
South Asia, constituting India, Pakistan, Bangladesh, Nepal, Sri Lanka and Bhutan represents the most populous region in the world burdened with significant multilevel poverty. Primary care is well-documented to reduce the burden of diseases (both chronic and acute) and associated morbidity and mortality; be cost-effective and improve health outcomes and health equity for both individuals and populations. Any field, for it to stay relevant and to effectively achieve its stated mission, needs to produce its own knowledge through research. This translates into patient care, practice models, education, and advocacy for health systems strengthening and reform. In primary care, weaving experiential wisdom with clinical evidence lies at the heart of translation. In this editorial, we highlight: a) that local context, in terms of geography, community, culture, language and beliefs, influences the clinical context and practice and therefore research. Thus, relying on primary care research done elsewhere in the globe will not suffice; b) effective primary care is based on a generalist approach that involves certain ways of being, knowing, perceiving, and doing guided by the values of humility and intellectual curiosity, diversity and inclusion, equity, holistic approach to evidence, integrity, transparency, accountability and adaptability, and communication; and c) that the South Asian Region (SAR) has a number of groups experimenting and innovating with various healthcare delivery systems, much of which is not known to the larger field. The WONCA SAR Primary Care Research Network has been set up to facilitate and support scholarship, writing, and publication in primary care especially by practising family physicians. In conclusion, it will be critical to simultaneously promote grounded theory research that integrates both the family medicine/primary care perspective and the voices of families and address the 'hidden curriculum' that shapes the attitudes and aspirations of young doctors at both the undergraduate and postgraduate level. Family Physicians treat a variety of patients on a daily basis, in order to improve the quality of care and impact of primary care, it is imperative that the understanding and application of research is enhanced by specialists of this discipline.


Hormonal control of a carbohydrate epitope involved in implantation in mice.
Ovariectomy and hormone replacement of mice were used to examine the hormonal control of expression on the uterine surface of a carbohydrate determinant (lacto-N-fucopentaose I, LNF I), involved in the initial interaction between the blastocyst and the endometrial epithelium at implantation. Pseudopregnant mice mated with sterile males were also used to elucidate the impact of embryonic signals on the expression of this antigen on the uterine surface. Two groups of fucosylated structures could be distinguished; one group was predominantly dependent on maternal oestrogen and progesterone, while the other group appeared to be less influenced by the hormonal milieu.


Coadministration of Trametinib and Palbociclib Radiosensitizes KRAS-Mutant Non-Small Cell Lung Cancers In Vitro and In Vivo.
To investigate the potential roles that p16 (CDKN2A) and RB activation have in sensitization to MEK inhibitor in resistant KRAS-mutant non-small cell lung cancer cells (NSCLC) in vitro and in vivo. Cell viability was measured with MTS assays. Effects of administration of radiation and combination drug treatments were evaluated by clonogenic assay, flow cytometry, and Western blots. DNA repair was assessed using immunofluorescent analysis. Finally, lung cancer xenografts were used to examine in vivo effects of drug treatment and radiation therapy. In this study, we showed that sensitivity to MEK inhibitor correlated to the RB/p16/CDK4 pathway and knockdown of RB induced resistance in cell lines sensitive to MEK inhibitor. Also, overexpression of p16 and inhibition of CDK4 had the ability to sensitize normally resistant cell lines. Our data indicated that the MEK inhibitor (trametinib, GSK112012) cooperated with the CDK4/6 inhibitor (palbociclib, PD0332991) to strongly reduce cell viability of KRAS-mutant NSCLCs that were resistant to the MEK inhibitor in vitro and in vivo. In addition, we report for the first time that resistance of KRAS-mutant NSCLCs to MEK inhibitor is, at least partly, due to p16 mutation status, and we described a drug combination that efficiently reactivates the RB tumor suppressor pathway to trigger radiosensitizing effects, apoptosis, and cell-cycle arrest. Our findings suggest that MEK inhibitor in combination with CDK4/6 inhibitor has significant anti-KRAS-mutant NSCLC activity and radiosensitizing effect in preclinical models, potentially providing a novel therapeutic strategy for patients with advanced KRAS-mutant NSCLCs.


There Is Still Room for Improvement: Presentation of a Neutral Borosilicate Glass with Improved Chemical Stability for Parenteral Packaging.
For pharmaceutical parenteral packaging the glass compositions have always been either Type I borosilicate or Type III soda-lime glass. As both the compositions and certain chemical and physical properties are mandated by international standards, there has not been room for any changes. However, by applying only minor adjustments, a borosilicate glass was developed that showed improved chemical stability. The chemical composition is still in the range of currently used borosilicate glasses, which makes it a Type I glass according to all current pharmacopeia. A study was performed on glass vials comparing the new glass with the standard FIOLAX(®) and two other publicly available glasses. In an extraction study with water at 121 °C the new glass showed the highest chemical stability with the lowest amount of extractables. In an accelerated ageing study, which was done with water, phosphate, and carbonate buffer at 40 °C for 12 months, the new glass also proved to have the lowest amount of leachables. In this article the new glass and the results from the studies are presented, showing the reader how much of an effect can be attained with only minor adjustments if the scientific fundamentals are clear. The pharmaceutical market has been quite constant and risk-oriented due to the high impact on the safety of the patient. As any change necessitates a complicated change process, this has, in consequence, lead the industry to resist changing the parenteral primary packaging material for decades. The main glasses have either been Type I borosilicate or Type III soda-lime glass. On the other hand, a combination of improved inspection systems and the development of more sensitive biologically based drugs has elevated the standards for parental packaging materials. For example, the measurement of extractables and leachables from the packaging material steadily came into focus. In this article, a new glass is presented that still belongs to the group of Type I borosilicate glasses according to all pharmacopeia. However, with some minor adjustments in the chemical composition it was possible to increase the chemical stability measurably. To prove this several studies were performed, of which the extraction study with water at 121 °C and the accelerated ageing study with water, phosphate, and carbonate buffer at 40 °C for 12 months are presented here.


The enhancement of hippocampal primed burst potentiation by dehydroepiandrosterone sulfate (DHEAS) is blocked by psychological stress.
This series of studies investigated the effects of psychological stress and the neurosteroid dehydroepiandrosterone sulfate (DHEAS) on hippocampal primed burst (PB) and long-term (LTP) potentiation, two electrophysiological models of memory. The DHEAS and stress manipulations were performed on awake rats, and then PB and LTP were recorded while the rats were anesthetized. DHEAS enhanced PB potentiation when administered to rats under non-stress conditions, but had no effect when given to stressed rats. Further study showed that DHEAS enhanced PB potentiation only when it was administered before, but not after, the rats were stressed. The DHEAS and stress manipulations had no effect on LTP. This study provides three major findings regarding stress, neurosteroids and hippocampal plasticity. First, DHEAS enhanced a threshold form of plasticity (PB potentiation), but had no effect on a supra-threshold form of plasticity (LTP). Second, stress blocked the DHEAS-induced enhancement of PB potentiation. Third, stress and DHEAS effects on the hippocampus were so durable they could be performed on awake animals and then be studied while the animals were anesthetized. That DHEAS enhanced a subset of forms of hippocampal plasticity under restricted behavioral conditions may help to resolve conflicting observations of DHEAS effects on cognition and mood in people.


[Risk of repeated hospitalizations in elderly users of an academic health center].
The aim of this study was to stratify 305 elderly (> 65 years) treated at an academic health center (Botucatu, São Paulo, Brazil) according to risk of repeated hospitalization. Data collection used an instrument to evaluate risk of repeated hospital admissions, including eight health indicators: self-rated health, hospitalizations, doctor visits, diabetes, cardiovascular disease, gender, social support, and age. 56.4% of interviewees presented low probability of repeated hospitalization, as compared to 26.9% medium, 10.5% medium-high, and 6.2% high probability. Combining health indicators with the probability of repeated hospitalization, for elderly classified as medium, medium-high, and high risk compared to low, the relative risk was significant: medium or bad health (2.31); hospitalization (2.38); > 3 doctor visits (1.75); diabetes (2.10); cardiovascular disease (2.76); male gender (1.68); and > 75 years of age (1.62). The instrument was able to stratify elderly at risk of repeated hospitalization and is thus useful for reorganizing public health services.


Clinical significance of Staphylococcus aureus bacteriuria at a tertiary care hospital.
Staphylococcus aureus bacteriuria has been associated with invasive S. aureus disease. The current project describes the clinical significance of S. aureus bacteriuria. We conducted a retrospective chart review of patients who had S. aureus bacteriuria at the Michael E. DeBakey Veterans Affairs Medical Center, 2008-2010. A total of 326 patients were included. Invasive S. aureus disease was documented within 12 months of bacteriuria in 56 patients (22.3% of methicillin-resistant S. aureus (MRSA) cases and 8.4% of methicillin-sensitive S. aureus (MSSA), p = 0.002). Multiple logistic regression indicated that MRSA bacteriuria (odds ratio (OR) 2.91, p = 0.010), absence of symptoms suggestive of a urinary tract infection (UTI) (OR 3.21, p = 0.019), inpatient status (OR 4.72, p = 0.003), and receipt of antibiotics active against S. aureus (OR 6.41, p < 0.001) were significantly associated with the presence of invasive S. aureus disease. Seventy-seven patients (23.6%) died within 12 months of the documented S. aureus bacteriuria. Age (OR 1.02, p = 0.025), absence of pyuria (OR 2.00, p = 0.029), the presence of invasive S. aureus disease (OR 2.05, p = 0.033), and inpatient status (OR 3.62, p < 0.001) were significantly associated with death. S. aureus bacteriuria is associated with significant morbidity and mortality. Patients without UTI symptoms, those with MRSA bacteriuria, and those without pyuria were more to likely to have worse outcomes (invasive S. aureus disease or death). Obtaining blood cultures should be considered in these cases.


A new force profile signal for a convex solution of muscle force estimation from electromyographic signals.
High-Density Surface Electromyography (HD-sEMG) is a non-invasive technique for measuring the electrical activity of a muscle with multiple, closely spaced electrodes. Estimation of muscle force is one of the applications of HD-sEMG. Usually, validating different EMG-Force models entails simple movements limited to laboratory settings. The validity of these models in more ecological conditions, requesting force production over a wide frequency band, remains unknown. In this study, we, therefore, compare the results of force prediction using four different types of input force profiles that can be representative of daily life activities, and we investigate whether the crest factor of these different input signals affects force prediction. For predicting the force from sEMG signals, we used our real-time and convex methods. HD-sEMG signals were recorded with 144 channels from the biceps brachii, brachioradialis, and triceps (long, lateral, and medial head) muscles of 24 healthy subjects during random signal, random phase, Schroeder phase, and minimum crest factor (crestmin) signal. The correlation and coefficient of determination (R2) between measured and predicted forces were calculated for the different force feedback profiles. The crestmin signal showed significantly better results based on statistical tests (P-value < 0.05), with correlation and R2 equal to 0.92±0.03 and 0.86±0.05, respectively. The results demonstrate that the crest factor of input signals is a crucial parameter that can impact the performance of EMG-Force models and must be considered during training.Clinical Relevance- This study demonstrates that lower crest factor multisine force profiles result in improved fitness for force prediction and can be used as an alternative to random signals.


Limited efficacy of uterine artery embolization for cervical leiomyomas.
To explore the effectiveness of uterine artery embolization (UAE) in treating symptomatic fibroids in the uterine cervix. Among 537 patients who underwent UAE, 10 who had fibroids located in the cervix were retrospectively analyzed. The mean diameter of the fibroids was 6.0 cm. Seven of the 10 patients presented a total of 10 fibroids in the uterine body or fundus simultaneously. Fibroids of the cervix and fibroids in the body or fundus were compared in terms of the effects of UAE on the treatment thereof and vascularity on angiographic findings. Cervical leiomyomas were classified into three grades based on the vascularity seen on aortography, from grade I, indicating poor vascularity, to grade III, indicating hypervascularity. Necrosis of fibroids was assessed by magnetic resonance imaging 3 months after UAE. Complete necrosis of leiomyomas in the uterine cervix was seen in only two of the 10 patients (20%), whereas all fibroids in the uterine body or fundus were completely infarcted (P < .05). Partial necrosis (PN) of the fibroid with a thin viable rim was seen in two patients, whereas PN with a thick rim was seen in four and no necrosis was seen in two. Grade I (ie, poor) vascularity was noted in five of nine patients (55.6%) with cervical fibroids larger than 3 cm. Poor vascularity was a frequent finding among cervical leiomyomas, and the outcomes of UAE for cervical leiomyomas were disappointing, indicating a need for caution in selecting and counseling patients for this treatment.


Central and peripheral cytokines mediate immune-brain connectivity.
The immune system is a homeostatic system that contributes to maintain the constancy of the molecular and cellular components of the organism. Immune cells can detect the intrusion of foreign antigens or alteration of self-components and send information to the central nervous system (CNS) about this kind of perturbations, acting as a receptor sensorial organ. The brain can respond to such signals by emitting neuro/endocrine signals capable of affecting immune reactivity. Thus, the immune system, as other physiologic systems, is under brain control. Under disease conditions, when priorities for survival change, the immune system can, within defined limits, reset brain-integrated neuro-endocrine mechanisms in order to favour immune processes at the expenses of other physiologic systems. In addition, some cytokines initially conceived as immune products, such as IL-1 and IL-6, are also produced in the "healthy" brain by glial cells and even by some neurons. These and other cytokines have the capacity to affect synaptic plasticity acting as mediators of interactions between astrocytes and pre- and post-synaptic neurons that constitute what is actually defined as a tripartite synapse. Since the production of cytokines in the brain is affected by peripheral immune and central neural signals, it is conceivable that tripartite synapses can, in turn, serve as a relay system in immune-CNS communication.


[Audio Computer-Assisted Interview: a new technology in the assessment of sexually transmitted diseases, HIV, and drug use].
Reliable responses are crucial when applying questionnaires on sensitive and stigmatized behaviors. This challenge has motivated researchers to develop new data collection methods. We conducted a systematic literature review on the implementation, effectiveness, reliability, and validity studies of ACASI (Audio Computer-Assisted Interview) in the assessment of drug use and sexual behavior. We reviewed 24 papers, none of which published by Brazilian researchers, and only three of which describing research implemented outside the United States. The studies showed that the computerized method is able to reduce psychological barriers linked to the collection of sensitive health-related information, thereby increasing its reliability. According to the surveys, the ACASI format is a fast and valid assessment method for large samples. ACASI generates databanks that can be easily processed and analyzed.


Misdiagnosed iuxta-articular osteoid osteoma of the calcaneus following an injury.
The diagnosis of osteoid osteoma, in usual localizations, is generally simple. In iuxta-articular localizations this tumor may be unrecognized and the diagnosis delayed. Injury has been sometimes correlated with the onset of symptoms and this can make the diagnosis even more difficult. We present a case of osteoid osteoma of the calcaneus iuxta-articular to the subtalar joint in a 17-year-old basketball player. He had a history of initial injury, ankle sprain during training, followed by pain and swelling. He was initially treated for lateral ligament lesion of the ankle with unsatisfactory results. After acute trauma the pain changed becoming chronic and mostly nocturnal disappearing when rofecoxib was taken. Standard x-ray didn't show the lesion. Nuclear Magnetic Resonance (NMR) and scintigraphic results were not well interpreted but these clinical changes and Computed Tomography (CT) images supported the diagnosis of osteoid osteoma. The complete resection of the bone lesion resolved all the symptoms and the histological report confirmed the suspected diagnosis.


A comparison of nuclear and mitochondrial cline shapes in a hybrid zone in the Sceloporus grammicus complex (Squamata; Phrynosomatidae).
The F5 and FM2 chromosome races of the Sceloporus grammicus complex form a hybrid zone in the Mexican state of Hidalgo. Previous studies of this zone have assessed genetic structure by averaging estimates of shape and width across three diagnostic chromosome markers. This approach is likely to mask subtle differences in cline shape among loci (e.g. selected vs. neutral), and obscure any displacement of cline centres (if present). Here we use maximum likelihood methods to construct the best fitting individual clines for three chromosomal markers, and also add two new markers; the mitochondrial DNA (mtDNA) locus, and the nuclear ribosomal DNA (rDNA) repeat. For each locus, hybrid zone models were fitted by cline shape and width, and the position and number of segments describing the centre of the zone. Pairwise comparisons between all clines revealed concordance between chromosomes 2 and 6, but significant discordance in cline structure among all other paired combinations. The concordance of chromosomes 2 and 6 suggests that these clines are maintained by genome-wide forces. The discordance of the chromosome 1 cline suggests an influence of asymmetric introgression, while the mtDNA cline is probably influenced by selection and drift. The rDNA locus reveals a pattern best explained by either extreme asymmetric introgression or gene conversion. The structure of zone indicates that genome-wide processes and locus specific selective forces as well as drift, are operating to different degrees on different loci. The locus-by-locus approach used here permits a finer discrimination among possible mechanisms responsible for the maintenance of the individual clines.


N-ethylmaleimide-modified subfragment-1 and heavy meromyosin inhibit reactivated contraction in motile models of retinal cones.
The mechanism of contraction in motile models of teleost retinal cones has been examined by using N-ethylmaleimide (NEM)-modified myosin fragments (NEM-S-1 and NEM-heavy meromyosin [HMM]) to prevent access of native myosin to actin filaments during reactivation of contraction. In the diurnal light/dark cycle, retinal cones of green sunfish (Lepomis cyanellus) and bluegill (lepomis macrochirus) exhibit length changes of more than 90 mum. The motile myoid region of the cone contracts from 100 mum in the dark to 6 mum in the light. Motile models for cone contraction have been obtained by lysis of dark-adapted retinas with the non-ionic detergent, Brij-58. These cone motile models undergo Ca(++)-and ATP-dependent reactivated contraction, with morphology and rate comparable to those observed in vivo (Burnside, B.,B. Smith, M. Nagata, and K. Porrello, 1982, J. Cell Biol., 92:198-206). The cone myoids contain longitudinally oriented actin filaments which bind myosin subfragment-1 (S-1) to form characteristic "arrowhead" complexes which dissociate in the presence of MgATP (Burnside, B., 1978, J. Cell Biol., 78:227-246). Modification of S-1 or HMM with the sulfhydryl reagent, NEM, produces new species, NEM-S-1 or NEM-HMM, which still bind actin but which fail to detach in the presence of MgATP (Meeusen, R.L., and W.Z. Cande, 1979, J. Cell Biol., 82:57-65). We have used NEM-S-1 and NEM-HMM to test whether cone contraction depends on an actomyosin force- generating system. We find that reactivated contraction of cone models is inhibited by NEM-S-1 and NEM-HMM but not by the unmodified species, S-1 and HMM. Thus, reactivated cone contraction exhibits NEM-S-1 and NEM-HMM sensitivity as well as Ca(++)- and ATP- dependence. These observations are consistent with and actimyosin-mediated mechanism for force production during cone contraction.


Sex, stress and their influence on respiratory regulation.
Much like locomotion or micturition, respiration is a rhythmic and stereotyped motor pattern controlled mainly by non-cortical structures including a complex circuit in the brainstem. Because tight regulation of lung ventilation is essential from the beginning of life, it has been presumed that the neural system regulating breathing is fixed, following a genetically predetermined developmental pattern. Here, we review evidence indicating that early life exposure to a non-systemic stress in the form of neonatal maternal separation (NMS) is sufficient to exert sex-specific consequences on the developmental trajectory of this vital homeostatic system that persist well into full maturity. At adulthood, male rats subjected to NMS are hypertensive and show an abnormally high hypoxic chemoreflex that correlates positively with respiratory instability during sleep. The effects are not observed in females. Investigation of the mechanisms this respiratory phenotype have highlighted the importance of 1) neuroendocrine influences on respiratory regulation and 2) stress-related imbalance between inhibitory (GABAergic) and excitatory (glutamatergic) modulation of the neural elements regulating breathing. These results provide new and valuable insight into the origins of respiratory disorders related to neural control dysfunction such as sleep disordered breathing.


Linguistic Predictors of Cultural Identification in Bilinguals.
Most of the world's population has knowledge of at least two languages. Many of these bilinguals are also exposed to and identify with at least two cultures. Because language knowledge enables participation in cultural practices and expression of cultural beliefs, bilingual experience and cultural identity are interconnected. However, the specific links between bilingualism and cultural identity remain largely unidentified. The current study examined which aspects of bilingualism relate to identification with first- and second-language cultures. Two-hundred-and-nine bilinguals completed a questionnaire probing linguistic background and cultural affiliations. Regression analyses indicated that cultural identification was predicted by age of language acquisition, language proficiency, foreign-accentedness, and contexts of long-term language immersion and current language exposure. Follow-up analyses revealed that the language-culture relations were mediated by the age and manner in which the second language was acquired. These findings are situated within a proposed framework of bilingual cultural identity. By identifying features of bilingualism that are relevant for cultural identity, the current research increases our understanding of the relationship between language and culture.


A Phase I Study of TRK-250, a Novel siRNA-Based Oligonucleotide, in Patients with Idiopathic Pulmonary Fibrosis.
Purpose: TRK-250 is a novel single-stranded oligonucleotide carrying a human Transforming growth factor-beta 1-targeting siRNA motif tethered by two proline linkers. Nonclinical studies have shown that TRK-250 may have potency to prevent the progression of pulmonary fibrosis. Herein, a phase I study was conducted to investigate the safety and pharmacokinetics (PKs) of TRK-250 in patients with idiopathic pulmonary fibrosis (IPF). Method: In the phase I study, 34 IPF patients were partially randomized to receive a placebo or TRK-250 in 4 single doses of 2, 10, 30, and 60 mg or multiple rising doses of 10, 30, and 60 mg once per week for 4 weeks by oral inhalation. For both the single- and multiple-dose studies, the primary endpoint was safety, and the secondary endpoint was PKs. Result: In all IPF patients who orally inhaled TRK-250, no significant drug-related adverse events (AEs) were observed. The AEs were mild or moderate, except for one severe case with acute exacerbation. One of the more common AEs was coughing. One patient discontinued treatment before the last dose because of coughing. There were no medically important findings related to safety endpoints based on clinical laboratory data (clinical chemistry, hematology, or urinalysis), vital signs data, electrocardiogram data, physical examination findings, pulse oximetry data, spirometry data, or diffusing capacity of the lung for carbon monoxide data. All the bioanalytical results of PKs in the blood were below the lower limit of quantification. Conclusions: Both the single and multiple doses of TRK-250 were safe and well tolerated in this first study done in IPF patients. Furthermore, TRK-250 was not detected in the systemic circulation following inhalation, indicating low or virtually nonexistent systemic exposure. This study is registered at ClinicalTrials.gov with identifier number NCT03727802.


[Restorative treatment by means of short porous-surfaced implants].
to assess clinical efficiency of restorative treatment on short porous-surfaced implants. One hundred and fifty-five porous-surfaced 5, 7 and 9 mm implants were placed in 71 patients with partial teeth loss and significant alveolar bone loss. Only one implant failure was observed one month after placement. Other 154 implants were used as restoration abutments. One year after restorative treatment mean periotest rate was -5,3, mean radiologicaly confirmed marginal bone loss 0,31 mm. No treatment failure were revealed during 3 years of follow-up.


Investigating walking environments in and around assisted living facilities: a facility visit study.
This study explores assisted living residents' walking behaviors, locations where residents prefer to walk, and walking environments in and around assisted living facilities. Regular walking is beneficial to older adults' physical and psychological health. Yet frail older residents in assisted living are usually too sedentary to achieve these benefits. The physical environment plays an important role in promoting physical activity. However, there is little research exploring this relationship in assisted living settings. The researcher visited 34 assisted living facilities in a major Texas city. Methods included walk-through observation with the Assisted Living Facility Walking Environment Checklist, and interviews with administrators by open- and close-ended questions. The data from 26 facilities were analyzed using descriptive statistics (for quantitative data) and content analysis (for qualitative data). The results indicate that (a) residents were walking both indoors and outdoors for exercise or other purposes (e.g., going to destinations); (b) assisted living facility planning and design details-such as neighborhood sidewalk conditions, facility site selection, availability of seating, walking path configuration (e.g., looped/nonlooped path), amount of shading along the path, presence of handrails, existence of signage, etc.-may influence residents' walking behaviors; and (c) current assisted living facilities need improvement in all aspects to make their environments more walkable for residents. Findings of the study provide recommendations for assisted living facilities to improve the walkability of environments and to create environmental interventions to promote regular walking among their residents. This study also implies several directions for future research.


Morphological variation of genetically confirmed Alouatta Pigra × A. palliata hybrids from a natural hybrid zone in Tabasco, Mexico.
While hybridization has been reported for a large number of primate taxa, there is a general lack of data on hybrid morphology for wild individuals with known genetic ancestry. A confirmed hybrid zone for the closely related Neotropical primates Alouatta palliata and A. pigra has provided a unique opportunity to study primate hybrid morphological variation. Here we used molecular evidence based on mitochondrial, Y-chromosome, and autosomal data to assess hybrid ancestry. We conducted univariate and multivariate statistical comparisons of morphometric data collected from individuals both outside and within the hybrid zone in Tabasco, Mexico. Our results show that of all the hybrids detected (N = 128), only 12% of them were approximately genetically intermediate, and none of them were first generation hybrids. Univariate pairwise comparisons among parental individuals, multigenerational backcrossed hybrids, and intermediate hybrids showed that overall, multigenerational backcrossed hybrids resemble the parental species with which they share most of their alleles. Conversely, intermediates were highly variable. Similarly, principal component analysis depicts an overlap between the parental species and their backcrosses when considering overall morphological differences. Finally, discriminant function analysis of the morphological variables was overall unreliable for classifying individuals into their assigned genotypic classes. Taken together, our results suggest that primate natural hybridization studies should incorporate molecular methods for determining ancestry, because morphology may not always be a reliable indicator of hybrid status. Hybrid zones could comprise a large number of multigenerational backcrossed hybrids that are indistinguishable from the parental species. The implications for studying hybridization in the primate fossil record are discussed.


Aberrant epigenetic regulation of GABRP associates with aggressive phenotype of ovarian cancer.
Metastasis is a major cause of therapeutic failure in ovarian cancer. To elucidate molecular mechanisms of ovarian cancer metastasis, we previously established a metastatic xenograft mouse model using human ovarian carcinoma SK-OV-3 cells. Using gene expression profiling, we found that γ-aminobutyric acid (GABA)A receptor π subunit (GABRP) expression was upregulated (>4-fold) in metastatic tissues from our xenograft mice compared with SK-OV-3 cells. Importantly, GABRP knockdown diminished the migration and invasion of SK-OV-3 cells, and reduced extracellular signal-regulated kinase (ERK) activation while overexpression of GABRP exhibited significantly increased cell migration, invasion and ERK activation. Moreover, treatment with the mitogen-activated protein kinase (MAPK)/ERK kinase (MEK) inhibitor U0126 similarly suppressed the migration and invasion of SK-OV-3 cells, implying that GABRP promotes these cellular behaviors by activating the MAPK/ERK pathway. Using genome-wide DNA methylation profiling, we identified hypomethylated CpG sites in the GABRP promoter in metastatic tissues from the xenograft mice compared with SK-OV-3 cells. Treatment with a DNA methyltransferase inhibitor demonstrated that methylation at -963 bp from the GABRP transcription start site (-963 CpG site) was critical for the epigenetic regulation of GABRP. Finally, we analyzed human ovarian cancer patient samples and showed DNA hypomethylation at the GABRP -963 CpG site in advanced stage, but not early-stage, primary tumors compared with their paired normal tissues. These findings suggest that GABRP enhances the aggressive phenotype of ovarian cancer cells, and that the DNA methylation status of the GABRP -963 CpG site may be useful for predicting the metastatic potential in ovarian cancer patients.


Mother-Infant Room-Sharing and Sleep Outcomes in the INSIGHT Study.
The American Academy of Pediatrics recommends infant-parent room-sharing until age 1. We assessed the association between room-sharing and sleep outcomes. The Intervention Nurses Start Infants Growing on Healthy Trajectories study is an obesity prevention trial comparing a responsive parenting intervention with a safety control among primiparous mother-infant dyads. Mothers completed the Brief Infant Sleep Questionnaire at 4, 9, 12, and 30 months. Reported sleep duration and overnight behaviors, adjusted for intervention group, were compared among early independent sleepers (own room <4 months), later independent sleepers (own room between 4 and 9 months), and room-sharers at 9 months. At 4 months, reported overnight sleep duration was similar between groups, but compared with room-sharers, early independent sleepers had better sleep consolidation (longest stretch: 46 more minutes, P = .02). At 9 months, early independent sleepers slept 40 more minutes nightly than room-sharers and 26 more minutes than later independent sleepers (P = .008). The longest stretch for early independent sleepers was 100 and 45 minutes more than room-sharers and later independent sleepers, respectively (P = .01). At 30 months, infants sleeping independently by 9 months slept >45 more minutes nightly than those room-sharing at 9 months (P = .004). Room-sharers had 4 times the odds of transitioning to bed-sharing overnight at both 4 and 9 months (P < .01 for both). Room-sharing at ages 4 and 9 months is associated with less nighttime sleep in both the short and long-term, reduced sleep consolidation, and unsafe sleep practices previously associated with sleep-related death.


The variant form of hairy-cell leukaemia.
Hairy-cell leukaemia-variant (HCL-variant) is a rare B-cell disorder which accounts for 10% of HCL cases. It affects elderly or middle-aged males. The main features are splenomegaly, lymphocytosis and cytopenias without monocytopenia. The circulating cells have a morphology intermediate between prolymphocytes and hairy cells. The immunophenotype shows a mature B-cell phenotype with expression of the B-cell antigens CD11c and CD103-but unlike typical HCL the cells are CD25- and HC2-negative. The histology of bone marrow and spleen shows a pattern of infiltration similar to that in HCL. There is no recurrent chromosomal abnormality but complex karyotypes and monoallelic p53 deletion by fluorescence in situ hybridization are common. Patients are resistant to alkylating agents and interferon-alpha (IFN-alpha) and only half achieve partial responses to pentostatin and/or cladribine. Splenectomy results in long-lasting partial responses in over two-thirds of the patients and is a good palliative treatment. Despite the lack of response to most therapies, the clinical course of HCL-variant is chronic. The median survival is 9 years and 42% of patients die of unrelated causes. Transformation to large cell is seen in 6% of patients. The inferior survival in HCL-variant compared with typical HCL cases may reflect the chemotherapy resistance.


Comparison of diffusion-weighted MR imaging and multidetector-row CT in the detection of liver metastases in patients operated for pancreatic cancer.
The purpose of this study is to determine the value of diffusion-weighted MR imaging (DWI) in the detection of liver metastases in patients with pancreatic tumors when compared to multidetector-row CT (MDCT). DWI and MDCT were performed in 31 consecutive patients with newly diagnosed, potentially resectable pancreatic tumors. CT images were obtained in the arterial and the portal venous phase. For DWI, a respiratory-triggered single-shot echo-planar imaging sequence (b values: 0, 300, and 600 s/mm(2)) was acquired. Images were analyzed in consensus by two radiologists blinded to the clinical data. Imaging results were correlated with intraoperative surgical and ultrasound findings as well as with results of histopathologic analysis and imaging follow-up. Sensitivity and specificity in detecting liver metastases were 53.3% and 77.8% for MDCT and 86.7% and 97.5% for DWI, respectively. In our study population DWI would have changed the therapeutic management in 4 out of 31 patients (12.9%) when compared to MDCT. In the present pilot study, DWI performed significantly better than MDCT in the detection of liver metastases in patients with pancreatic tumors. Therefore, DWI may help to optimize therapeutic management in those patients in the future.


Serum-mediated suppression of lymphocyte transformation responses in coccidioidomycosis.
Lymphocyte transformation (LT) responses to coccidioidin (CDN) and spherulin were suppressed in 11 (73%) of 15 patients with active coccidioidomycosis when their mononuclear cells were assayed in autologous serum as compared to serum from healthy, CDN skin test-positive subjects. Suppressed LT responses were specific for Coccidioides immitis antigens in 7 (64%) of the 11 patients. Immunoaffinity chromatography of patient sera with Staphylococcus protein A adsorbed the suppressor component(s) and thereby established that suppression was attributed to immunoglobulin G, either alone or complexed with antigen. The possibility that suppression was mediated by immune complexes was examined by adding complexes formed in vivo or in vitro to mononuclear cell cultures of healthy CDN-reactive persons before LT assays. Although complexes prepared in this manner were reactive in an enzyme-linked immunosorbent assay designed to detect Coccidioides antigen-specific immune complexes, no suppression of LT responses was observed. We conclude that serum-mediated suppression of LT responses in coccidioidomycosis is attributed to monomeric and not immune-complexed immunoglobulin G antibody.


[Noise generated during incubator manipulation: implications for nursing care].
Uncontrolled experimental study. Quantify the impact noise of the 23 incubators functioning in the intensive and intermediate care units of a public university hospital in Ribeirão Preto-SP. Measuring occurred during incubator manipulation in a careful and in a rough way, considering the impact noise generated by the various situations in which the incubator was manipulated. In practically all manipulation situations in both the careful and the rough way, group B presented higher NPS, followed by group A and D. Group C demonstrated the lowest NPS during the different manipulation situations. Even careful manipulation generated a strong noise level, and NPS almost always doubled or tripled when the same situation was realized roughly. Careful manipulation, together with lower incubator use time, revealed to be effective in reducing the noise produced during equipment manipulation.


[Ischemic type of diabetic maculopathy].
The pathogenesis of diabetic maculopathy is discussed. The possibility of different diagnostic methodic (OCT, FAG and etc.) is shown. Positive results of selective coagulation of pigment epithelium are achieved.


The Association between BZIP Transcription Factors and Flower Development in Litsea cubeba.
The basic leucine zipper (bZIP) family is one of the largest families of transcription factors among eukaryotic organisms. Members of the bZIP family play various roles in regulating the intricate process of flower development in plants. Litsea cubeba (Lour.) (family: Lauraceae) is an aromatic, dioecious plant used in China for a wide range of applications. However, no study to date has undertaken a comprehensive analysis of the bZIP gene family in L. cubeba. In this work, we identified 68 members of the bZIP gene family in L. cubeba and classified them into 12 subfamilies based on previous studies on Arabidopsis thaliana. Transcriptome data analysis revealed that multiple LcbZIP genes exhibit significantly high expression levels in the flowers of L. cubeba, while some also demonstrate distinct temporal specificity during L. cubeba flower development. In particular, some LcbZIP genes displayed specific and high expression levels during the stamen and pistil degradation process. Using differential gene expression analysis, weighted gene co-expression network analysis, and Gene Ontology enrichment analysis, we identified six candidate LcbZIP genes that potentially regulate stamen or pistil degradation during flower development. In summary, our findings provide a framework for future functional analysis of the LcbZIP gene family in L. cubeba and offer novel insights for investigating the mechanism underlying pistil and stamen degeneration in this plant.


Use of bronchodilators during non-invasive mechanical ventilation.
Bronchodilators represent one of the most important therapeutic weapons for the treatment of airway obstructive diseases and the inhaled route of administration is very often employed due to the greater drug availability and reduced magnitude of side effects. During acute exhacerbations, it is not unfrequent that the elastic and resistive loads imposed on the ventilatory pump overcome the force sustainable by the respiratory muscles and the patient requires ventilatory assistance, in order to relieve fatigue and to optimize alveolar gas exchange. During these episodes, inhaled bronchodilators, far from being discontinued, sometime must be administered during mechanical ventilation, that, in hypercapnic ventilatory failure can be frequently applied noninvasively with a good rate of success. While in the current literature there are a lot of data about inhaled drug administration during invasive mechanical ventilation, very few data are available on the topic of aerosol therapy during noninvasive mechanical ventilation. With the present paper we want to analyze the rationale, the feasibility and the current data dealing with the administration of inhaled drugs during noninvasive mechanical ventilation.


Technological innovations in shock wave lithotripsy.
Since 1980, extracorporeal shock wave lithotripsy (SWL) has been employed in the treatment of urolithiasis, offering noninvasive alternatives to surgical techniques. In addition to being limited by the size and location of the stones, its efficacy is influenced by several factors. Despite the advancement of other surgical techniques, SWL could maintain its position with new improvements. Our objective is to review the existing literature on the latest advances in the extracorporeal treatment of lithiasis. A non-systematic literature review was carried out from 2017 to 2023 to obtain 26 articles on three different emerging technologies in extracorporeal lithotripsy: Burst Wave Lithotripsy (BWL), Histotripsy, and Microbubble Lithotripsy (ML). The BWL uses sinusoidal bursts of US waves delivered at lower and higher frequencies than conventional SWL. Its mechanism of action generates a higher quality fragmentation (fine fragments) instead of generating tensile stresses for stone fracture resulting in larger fragments, as in traditional SWL. Studies in pigs and humans have shown effective fragmentation with a good safety profile. Based on High Intensity Focused Ultrasound (HIFU) technology, histotripsy fragments tissue through cavitation. Good in vitro results have been shown, but the formation of microbubbles between the stone and ultrasound waves hinders the progress of this technique. Microbubble Lithotripsy (ML) combines microbubbles and ultrasound for safe and effective stone fragmentation. In vitro and pig results are promising. This technique can help optimize treatments and reduce energy levels. Technological innovation is not only being applied to endourological techniques, but also to ESWL. New techniques such as BWL, histotripsy and ML are promising, with good results in the research phase.


4-Amino-2-Sulfanylbenzoic Acid as a Potent Subclass B3 Metallo-β-Lactamase-Specific Inhibitor Applicable for Distinguishing Metallo-β-Lactamase Subclasses.
The number of cases of infection with carbapenem-resistant Enterobacteriaceae (CRE) has been increasing and has become a major clinical and public health concern. Production of metallo-β-lactamases (MBLs) is one of the principal carbapenem resistance mechanisms in CRE. Therefore, developing MBL inhibitors is a promising strategy to overcome the problems of carbapenem resistance conferred by MBLs. To date, the development and evaluation of MBL inhibitors have focused on subclass B1 MBLs but not on B3 MBLs. In the present study, we searched for B3 MBL (specifically, SMB-1) inhibitors and found thiosalicylic acid (TSA) to be a potent inhibitor of B3 SMB-1 MBL (50% inhibitory concentration [IC50], 0.95 μM). TSA inhibited the purified SMB-1 to a considerable degree but was not active against Escherichia coli cells producing SMB-1, as the meropenem (MEM) MIC for the SMB-1 producer was only slightly reduced with TSA. We then introduced a primary amine to TSA and synthesized 4-amino-2-sulfanylbenzoic acid (ASB), which substantially reduced the MEM MICs for SMB-1 producers. X-ray crystallographic analyses revealed that ASB binds to the two zinc ions, Ser221, and Thr223 at the active site of SMB-1. These are ubiquitously conserved residues across clinically relevant B3 MBLs. ASB also significantly inhibited other B3 MBLs, including AIM-1, LMB-1, and L1. Therefore, the characterization of ASB provides a starting point for the development of optimum B3 MBL inhibitors.


Toward Successful Implementation of Speech Recognition Technology: A Survey of SRT Utilization Issues in Healthcare Settings.
To evaluate physician utilization of speech recognition technology (SRT) for medical documentation in two hospitals. A quantitative survey was used to collect data in the areas of practice, electronic equipment used for documentation, documentation created after providing care, and overall thoughts about and satisfaction with the SRT. The survey sample was from one rural and one urban facility in central Missouri. In addition, qualitative interviews were conducted with a chief medical officer and a physician champion regarding implementation issues, training, choice of SRT, and outcomes from their perspective. Seventy-one (60%) of the anticipated 125 surveys were returned. A total of 16 (23%) participants were practicing in internal medicine and 9 (13%) were practicing in family medicine. Fifty-six (79%) participants used a desktop and 14 (20%) used a laptop (2%) computer. SRT products from Nuance were the dominant SRT used by 59 participants (83%). Windows operating systems (Microsoft, Redmond, WA) was used by more than 58 (82%) of the survey respondents. With regard to user experience, 42 (59%) participants experienced spelling and grammatical errors, 15 (21%) encountered clinical inaccuracy, 9 (13%) experienced word substitution, and 4 (6%) experienced misleading medical information. This study shows critical issues of inconsistency, unreliability, and dissatisfaction in the functionality and usability of SRT. This merits further attention to improve the functionality and usability of SRT for better adoption within varying healthcare settings.


The endogenous modulation of IOR is nasal-temporal asymmetric.
Inhibition of return (IOR) refers to a reflexive mechanism mediated by phylogenetically primitive extrageniculate visuomotor pathways, which apparently serves to favor novel spatial locations by inhibiting those recently sampled. We demonstrate an asymmetry between temporal and nasal hemifields in the strategic modulation of IOR by endogenously controlled attention. Exogenous and endogenous precues were manipulated independently on each trial such that precues to initiate endogenous spatial orienting were presented after IOR had been activated by exogenous visual signals. Both types of precues manifested their characteristic effects on reaction time (RT) to detect subsequent targets: facilitation by endogenous precues, and IOR by exogenous precues. Under monocular viewing, an asymmetric interaction between these two mechanisms was observed. While endogenous allocation of attention to the nasal hemifield reduced IOR, no endogenous modulation of IOR was present in the temporal hemifield where the effects of the two types of precues were independent. These observations suggest a framework for understanding the neurobiology of automaticity and control - from an evolutionary perspective.


Myelin proteolipid protein gene structure and its regulation of expression in normal and jimpy mutant mice.
The mouse proteolipid protein (PLP) gene was cloned into the lambda bacteriophage Charon 4A. The organization and the nucleotide sequence of the exons of the mouse PLP gene were quite similar to those of their human counterparts, consisting of seven exons. The transcription of the PLP gene started from multiple sites. There was a unique sequence tandemly repeated four times, sharing homology with the herpes simplex virus DR2 sequence, upstream from the transcribed region. Expression of the myelin basic protein (MBP) is also restricted to the oligodendrocytes in the central nervous system as is the PLP expression. Homology search against the mouse MBP gene revealed that several boxes in the 5'-flanking region of PLP show a high degree of homology with the sequence present in the MBP 5'-flanking region, possibly of importance in the concomitant expression of both genes in the central nervous system. PLP-mRNA in jimpy mutant mice does not contain exon 5 and its content is greatly reduced. We analyzed the jimpy PLP-mRNA and showed that the transcription initiated from the same sites as those in normal mice. Cloning and sequencing of the 5'-flanking region of the jimpy PLP gene revealed that there were no mutations in the promoter region of the jimpy PLP gene. Therefore, it is likely that a mutation, presumably existing within the jimpy PLP gene, caused the skipping of exon 5 and directly affected the mRNA level.


Prevention of compartment syndrome associated with dorsal lithotomy position.
There is a clear association between the dorsal lithotomy position and the development of a postoperative compartment syndrome. Compartment syndrome occurs when elevated pressure in an osteofascial compartment compromises local perfusion and often results in neurovascular damage and permanent disability. Surgeons must recognize this association and provide appropriate preventative measures, have a high index of suspicion in all patients, and be prepared to make an early diagnosis when it is encountered. We will report four cases we encountered, discuss the etiology, analyze the literature, and propose a modification on patient positioning that has minimized the incidence of compartment syndrome at our institution. A variety of intraoperative events, including hypotension, hypoxemia, leg elevation, hip and knee flexion, direct pressure, and compressive bandages may all contribute to compartmental ischemia. Ischemia is followed by reperfusion, capillary leakage from the ischemic tissue, and a further increase in tissue edema. Increased edema causes further embarrassment of perfusion and perpetuation of the cycle, ultimately resulting in neuromuscular compromise secondary to ischemia. To prevent this destructive cycle, emphasis must be placed on maintaining a high index of suspicion in all patients, minimizing leg elevation, and on the importance of early diagnosis and intervention.


Human immunoglobulin M memory B cells controlling Streptococcus pneumoniae infections are generated in the spleen.
Splenectomized and asplenic patients have a high incidence of infections by encapsulated bacteria and do not respond to polysaccharide vaccines. To understand whether the absence of the spleen is associated with a defined B cell defect, we analyzed B cell subsets in the peripheral blood. We found that a population of B cells known as immunoglobulin (Ig)M memory is lacking in patients without spleen. The absence of IgM memory B cells correlates with an impaired immune response to encapsulated bacteria not only in splenectomized patients, but also in individuals with an intact spleen. We show that the physiological and transient predisposition to pneumococcal infections of young children (0-2 yr) is associated with the lack of circulating IgM memory B cells and of serum antipolysaccharide IgM. We also demonstrate that IgM memory B cells are undetectable in a fraction of patients with common variable immunodeficiency, who have recurrent and invasive infections by encapsulated bacteria. IgM memory B cells, therefore, require the spleen for their generation and/or survival and are responsible for the protection against encapsulated bacteria.


Monitoring walking and cycling of middle-aged to older community dwellers using wireless wearable accelerometers.
Changes in gait parameters have been shown to be an important indicator of several age-related cognitive and physical declines of older adults. In this paper we propose a method to monitor and analyze walking and cycling activities based on a triaxial accelerometer worn on one ankle. We use an algorithm that can (1) distinguish between static and dynamic functional activities, (2) detect walking and cycling events, (3) identify gait parameters, including step frequency, number of steps, number of walking periods, and total walking duration per day, and (4) evaluate cycling parameters, including cycling frequency, number of cycling periods, and total cycling duration. Our algorithm is evaluated against the triaxial accelerometer data obtained from a group of 297 middle-aged to older adults wearing an activity monitor on the right ankle for approximately one week while performing unconstrained daily activities in the home and community setting. The correlation coefficients between each of detected gait and cycling parameters on two weekdays are all statistically significant, ranging from 0.668 to 0.873. These results demonstrate good test-retest reliability of our method in monitoring walking and cycling activities and analyzing gait and cycling parameters. This algorithm is efficient and causal in time and thus implementable for real-time monitoring and feedback.


Cigarette smoking and chlorpromazine disposition and actions.
Chlorpromazine, 75 mg, was taken by eight cigarette smokers and nine nonsmokers. Mean maximum percent decrease in mean blood pressure with change in position from lying to standing during the first 10 hr after chlorpromazine was 46% greater in nonsmokers than in smokers. Of the eight smokers, six did not become sleepy after chlorpromazine, one became slightly sleepy, and one fell asleep. Of the nine nonsmokers, three became slightly sleepy, one moderately sleepy, and five fell asleep. Mean peak plasma concentration of chlorpromazine was 24% lower and mean area under the plasma concentration-time curve (AUC) was 36% lower in smokers than in nonsmokers. There was no correlation between these plasma level parameters and either the degree of sleepiness or the degree of orthostatic hypotension in the subjects. The relatively small differences between cigarette smokers and nonsmokers in AUCs and peak plasma levels for chlorpromazine, combined with the lack of relationship between these plasma level parameters, the occurrence of drowsiness, and the magnitude of orthostatic hypotension in our subjects make it unlikely that cigarette smoking influenced these effects of chlorpromazine by enhancing its overall metabolism.


[Acute abdomen for perforated gastrointestinal stromal tumor (GIST). A case report].
The authors report on a case of voluminous perforated gastrointestinal stromal tumor (GIST) of small intestine and make a review to the light of most recent clinicopathologic advancements. The first clinical manifestation as acute abdomen due to their perforation extremely rare. Gastrointestinal stromal tumor (GIST) represent a rare group of multiform tumors with various biological behaviour. Were identified in the past as leiomyomas, leiomyosarcomas or leiomyoblastomas, has been reclassified on immunochemical features, with a positive expression of Kit (CD117 antigen). Traditionally the prognostic factors of these tumour are: mitotic rate, tumor size and anatomic site. The complete surgical resection and use of imatinib mesylate (Gleevec) are the best available approach.