Nanoconfinement Effect for Signal Amplification in Electrochemical Analysis and Sensing.
Owing to the urgent need for electrochemical analysis and sensing of trace target molecules in various fields such as medical diagnosis, agriculture and food safety, and environmental monitoring, signal amplification is key to promoting analysis and sensing performance. The nanoconfinement effect, derived from nanoconfined spaces and interfaces with sizes approaching those of target molecules, has witnessed rapid development for ultra-sensitive analyzing and sensing. In this review, the two main types of nanoconfinement systems - confined nanochannels and planes - are assessed and recent progress is highlighted. The merits of each nanoconfinement system, the nanoconfinement effect mechanisms, and applications for electrochemical analysis and sensing are summarized and discussed. This review aims to help deepen the understanding of nanoconfinement devices and their effects in order to develop new analysis and sensing applications for researchers in various fields.


Photoaffinity labeling of mitochondrial adenosine triphosphatase by an azido derivative of the natural adenosine triphosphate inhibitor.
The natural mitochondrial ATPase inhibitor (IF1) was modified with a radioactivity labeled heterobifunctional and photosensitive reagent, methyl 4-azido(14C)benzimidate ((14C)MABI). Titration experiments of IF1 by (14C)MABI and tryptic maps of (14C)MABI-IF1 indicated that specific lysine residues in IF1 are preferentially labeled by (14C)MABI. Under appropriate conditions of labeling (1 to 2 lysine residues modified per IF1), MABI-IF1 exhibited the same inhibitory potency as native IF1 on the hydrolytic activity of the coupling factor 1 of mitochondrial ATPase (F1). The same conditions were required for inhibition of F1 by MABI-IF1 and IF1 (slightly acidic pH and presence of ATP and MgCl2). In photolabeling experiments, (14C)MABI-IF1 was used to investigate the localization of IF1 binding sites on F1. Upon photoirradiation, MABI-IF1 bound selectively to the beta subunit of soluble or membrane-bound F1. Adenylyl imidodiphosphate and quercetin, two compounds which partially mimic the inhibitory effect of IF1 on ATPase activity of F1, markedly prevented the binding of (14C)MABI-IF1 to F1; on the other hand, aurovertin, a specific ligand of the beta subunit of F1, did not affect the interaction between (14C)MABI-IF1 and F1. In the absence of light, (14C)MABI-IF1 was used as a reversible radiolabeled ligand with respect to membrane bound F1 to investigate F1-IF1 interactions to inside-out submitochondrial particles as a function of the energy state of the particles. Oxidation of NADH by submitochondrial particles resulted in a decrease of bound (14C)MABI-IF1; the effect was counteracted by antimycin. The data suggested that added (14C)MABI-IF1 is capable of exchanging with IF1 bound to F1 in submitochondrial particles and that the rate and extent of (14C)MABI-IF1 release are triggered by the proton-motive force developed by the particles.


Biofabrication of Human Skin with Its Appendages.
Much effort has been made to generate human skin organ in the laboratory. Yet, the current models are limited due to the lack of many critical biological and structural features of the skin. Importantly, these in vitro models lack appendages and fail to recapitulate the whole human skin construction. Thus, engineering a human skin with the capacity to generate all components, including appendages, is a major challenge. This review intends to provide an update on the recent efforts underway to regenerate appendage-bearing skin organs based on scaffold-free and scaffold-based bioengineering approaches. Although the mouse skin equivalents containing hair follicles, sebaceous glands, and sweat glands have been established in vitro, there has been limited success in humans. A combination of biofabricated matrices and cell aggregates, such as organoids, can pave the way for generating skin substitutes with human-like biological, structural, and physical features. Accordingly, the formation of human skin organoids and reconstruction of vascularized skin equipped with immune cells prompt calls for more scientific research. The generation of appendage-bearing skin substitutes can be applied in practice for wound healing, hair restoration, and scar treatment.


Social and Environmental Determinants of Community-Acquired Antimicrobial-Resistant Escherichia coli in Children Living in Semirural Communities of Quito, Ecuador.
Extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL), a family of bacteria that includes Escherichia coli, have emerged as a global health threat. This study examined risks associated with carriage of third-generation cephalosporin-resistant (3GC-R) E. coli, including ESBL-producing, multidrug-resistant, and extensively drug-resistant (XDR) strains in children living in semirural parishes of Quito, Ecuador. We conducted a longitudinal study with two cycles of sampling (N = 374, N = 366) that included an analysis of child fecal samples and survey questions relating to water, sanitation, and hygiene, socioeconomic status, household crowding, and animal ownership. We used multivariate regression models to assess risk factors associated with a child being colonized. Across the two cycles, 18.4% (n = 516) of the 3GC-R isolates were ESBL-producing E. coli, and 40.3% (n = 516) were XDR E. coli. Children living in households that owned between 11 and 20 backyard animals had an increased odds of being colonized with XDR E. coli (odds ratio [OR] = 1.94, 95% confidence interval [CI]: 1.05-3.60) compared with those with no animals. Households that reported smelling odors from commercial poultry had increased odds of having a child positive for XDR E. coli (OR = 1.72, 95% CI: 1.11-2.66). Our results suggest that colonization of children with antimicrobial-resistant E. coli is influenced by exposure to backyard and commercial livestock and poultry. Future studies should consider community-level risk factors because child exposures to drug-resistant bacteria are likely influenced by neighborhood and regional risk factors.


Development and application of a multiplex polymerase chain reaction for avian respiratory agents.
A multiplex polymerase chain reaction (PCR) was developed and optimized to simultaneously detect 6 avian respiratory pathogens. Six sets of specific oligonucleotide primers for infectious bronchitis virus (IBV), avian influenza virus (AIV), infectious laryngotracheitis virus (ILTV), Newcastle disease virus (NDV), Mycoplasma gallisepticum (MG), and Mycoplasma synoviae (MS) were used respectively in the test. With the use of agarose gel electrophoresis for detection of the PCR-amplified DNA products, the sensitivity of detection was between 10 pg for IBV, AIV, MG, and ILTV and 100 pg for NDV and MS after 35 cycles of PCR. Similar sensitivity of these primers was achieved with chickens experimentally infected with respiratory pathogens. In experimental infections, the multiplex PCR was able to detect all the infected chickens in each group at I and 2 wk postinfection as compared with serologic tests at 2 wk postinfection that confirmed the presence of specific antibodies. The multiplex PCR was also able to detect and differentiate coinfections with two or more pathogens. No specific DNA amplification for respiratory avian pathogens was observed among noninoculated birds kept separately as a negative control group.


Allergenicity of Maillard reaction products from peanut proteins.
It is known that peanut allergy is caused by peanut proteins. However, little is known about the impact of roasting on the allergenicity of peanuts. During roasting, proteins react with sugars to form Maillard reaction products, which could affect allergenicity. To determine if the Maillard reaction could convert a nonallergenic peanut protein into a potentially allergenic product, nonallergenic lectin was reacted with glucose or fructose at 50 degrees C for 28 days. Browning products from heat-treated peanuts were also examined. The products were analyzed in immunoblot and competitive assays, using a pooled serum (i.e., IgE antibodies) from patients with peanut anaphylaxis. Results showed that the products were recognized by IgE and had an inhibitory effect on IgE binding to a peanut allergen. Thus, the findings suggest that these Maillard reaction products are potentially allergenic and indicate the need to verify whether the Maillard reaction products formed in peanuts during roasting increase their allergenicity.


New York State urban and rural measurements of continuous PM2.5 mass by FDMS, TEOM, and BAM.
Field evaluations and comparisons of continuous fine particulate matter (PM2,5) mass measurement technologies at an urban and a rural site in New York state are performed. The continuous measurement technologies include the filter dynamics measurement system (FDMS) tapered element oscillating microbalance (TEOM) monitor, the stand-alone TEOM monitor (without the FDMS), and the beta attenuation monitor (BAM). These continuous measurement methods are also compared with 24-hr integrated filters collected and analyzed under the Federal Reference Method (FRM) protocol. The measurement sites are New York City (the borough of Queens) and Addison, a rural area of southwestern New York state. New York City data comparisons between the FDMS TEOM, BAM, and FRM are examined for bias and seasonality during a 2-yr period. Data comparisons for the FDMS TEOM and FRM from the Addison location are examined for the same 2-yr period. The BAM and FDMS measurements at Queens are highly correlated with each other and the FRM. The BAM and FDMS are very similar to each other in magnitude, and both are approximately 25% higher than the FRM filter measurements at this site. The FDMS at Addison measures approximately 9% more mass than the FRM. Mass reconstructions using the speciation trends network filter data are examined to provide insight as to the contribution of volatile species of PM2.5 in the FDMS mass measurement and the fraction that is likely lost in the FRM mass measurement. The reconstructed mass at Queens is systematically lower than the FDMS by approximately 10%.


Surgical site infections following caesarean sections at Emirati teaching hospital: Incidence and implicated factors.
The rate of delivery by caesarean sections is increasing globally and, therefore, the incidence of post-caesarean surgical site infections (SSIs) is probably also going to rise. The aim of the present study was to determine the incidence of SSIs after caesarean operations and to explore the factors associated with an increased risk of post-caesarean SSIs. A retrospective study was performed to assess all women who underwent caesarean sections from January 2016 to December 2017 at Al Ain Hospital in the United Arab Emirates (UAE). Backward multivariate logistic regression analysis was utilized to specify the variables that were significantly and independently connected with the development of post-caesarean SSIs. In total, 807 women underwent caesarean deliveries at the study site hospital during the two-year study period (January 2016-December 2017). Post-operative SSI was detected in 11 (1.4%) of the women who underwent caesarean operations. Of these, 11 (100%) women were diagnosed post-discharge, within 30 days after the date of the surgery. Multivariate logistic regression analysis showed that increased gestational age (P = 0.045) was significantly and independently associated with the development of post-caesarean SSI. Increased gestational age was found to be an independent predictor of post-caesarean SSIs. This identified risk factor should inform targeted health care policies to reduce the rate of SSIs.


Sex Differences in monocytes and TLR4 associated immune responses; implications for systemic lupus erythematosus (SLE).
It has been shown that TLR7 and TLR9 signaling play a role in SLE pathogenesis. Our recent study revealed that estrogen receptor α knockout mice have impaired inflammatory responses to TLR3, TLR4, TLR7 and TLR9 ligand stimulation in DCs, B cells and whole spleen cells. These findings indicate that estrogen receptor mediated signaling may impact universal TLR responsiveness. Whether estrogen has a direct or indirect effect on TLR responsiveness by immune cells is not clear. There is evidence of a role of TLR4 in SLE disease pathogenesis, such as the kidney damage, the induction of CD40 and autoantibodies, the suppression of regulatory T cells, and the role of pro-inflammatory cytokines (e.g., IL-6, IL-1β, TNF-α) in SLE pathogenesis that can be induced by TLR4-mediated monocyte activation, suggesting that TLR4 and TLR4 responsiveness are also important for SLE disease. This review will focus on TLR4 responses and monocytes, which are understudied in systemic autoimmune diseases such as SLE.


[Neighborhood characteristics and individual health under Korean context].
Recently much attention has been paid to the effect of neighborhood characteristics on the health of individuals, independent of individual demographic and/or socioeconomic characteristics. Although many empirical studies of a kind, mostly based on Western society, have appeared on various international journals, few studies have shown empirical evidence of neighborhood characteristics as an independent and significant risk factor of ill health in Korea. This paper discusses possible reasons that neighborhood seems to be neither significant nor substantial regarding its impact on the health of Koreans. Addressing the uniqueness of Korean society regarding the concept of neighborhood, attributes of neighborhoods, and methodological challenges, authors suggest that more sophisticated conceptual and methodological approach, specific to Korean society, should enable to reveal the effect of neighborhood characteristics on individual health in Korea.


Brain Cooling: An Economy Mode of Temperature Regulation in Artiodactyls.
Artiodactyls employ selective brain cooling (SBC) regularly during experimental hyperthermia. In free-ranging antelopes, however, SBC often was present when body temperature was low but absent when brain temperature was near 42 degrees C. The primary effect of SBC is to adjust the activity of the heat loss mechanisms to the magnitude of the heat stress rather than to the protection of the brain from thermal damage.


Arterial flow measurements during reactive hyperemia using NIRS.
Non-invasive evaluation of peripheral perfusion may be useful in many contexts including clinical research. We validated a novel non-invasive spectroscopy technique to quantify forearm arterial inflow. This method, which is based on the measurement of tissular total hemoglobin variations after an ischemic period, was compared to strain gauge plethysmography (SGP). The technique uses near-infrared spectroscopy (NIRS) to determine the rate of change of forearm tissue oxygenation during reactive hyperemia. In this study, 13 subjects were simultaneously evaluated with NIRS and SGP. Nine baseline flow measurements were performed to assess the reproducibility of each method. Twenty-seven serial measurements were then made to evaluate flow variation during forearm reactive hyperemia. SGP and NIRS methods showed excellent reproducibility with the same intra-class correlation coefficients (0.98). In conclusion, the NIRS technique appears well suited for non-invasive evaluation of quantitative arterial forearm flow.


Properties of calcium-dependent regulatory proteins from fungi and yeast.
Calmodulins were isolated from vegetative mycelia of Basidiomycetes fungi, Agaricus campestris and Coprinus lagopus. These calmodulins showed similar mobilities to those of animal calmodulins on nondenaturing polyacrylamide gel electrophoresis in the presence or absence of Ca2+. The molecular weights of both calmodulins were determined to be 16,000. Agaricus calmodulin consisted of 148 amino acids including epsilon-N-trimethyllysine and cysteine. The UV-absorption spectrum showed the relatively high content of phenylalanine in Basidiomycetes calmodulins. The difference UV-absorption spectrum due to the blue shift by Ca2+ was observed. Both calmodulins activated muscle myosin light chain kinase and pea NAD+ kinase in a Ca2+-dependent manner, and the activities were inhibited by trifluoperazine or chlorpromazine. A calmodulin-like protein was partially purified from baker's yeast, Saccharomyces cerevisiae. However, detection of a calmodulin-like protein in prokaryotes was not successful.


Activation of NF-kappa B and elevation of MnSOD gene expression by thiol reducing agents in lung adenocarcinoma (A549) cells.
The effect of reducing agents, including N-acetylcysteine (NAC), dithiothreitol (DTT), and 2-mercaptoethanol (2-ME) on nuclear transcription factor-kappa B (NF-kappa B) activation and manganese superoxide dismutase (MnSOD) expression was investigated in a pulmonary adenocarcinoma (A549) cell line. NAC, DTT, and 2-ME each activated the transcription factor NF-kappa B and increased steady-state levels of MnSOD mRNA and enzyme activity in these cells. In addition, NAC, DTT, and 2-ME increased chloramphenicol acetyltransferase (CAT) activity in cells transfected with a construct containing the CAT gene under the control of the rat MnSOD promoter. SOD and catalase (500 U/ml) plus ethanol (1 mM) did not inhibit activation of NF-kappa B or elevation of steady-state MnSOD mRNA levels by NAC, DTT, or 2-ME. Controls in which comparable amounts of O2-. to those produced by thiols were generated by hypoxanthine and xanthine oxidase, or in which H2O2 was added directly, had neither activated NF-kappa B nor elevated MnSOD mRNA. This shows that reactive oxygen intermediates, which may be formed during autooxidation, may not contribute to activation of NF-kappa B. Because the MnSOD promoter also contains potential binding sites for other transcription factors, such as promoter-selective transcription factor-1 (SP-1), activator protein-1 (AP-1), AP-2, adenosine 3',5'-cyclic monophosphate-regulator element binding factor (CREB), and transcription factor IID complex (TFIID), the effect of thiols on their activation also were evaluated. In contrast to findings with NF-kappa B, there was only minor activation of AP-1 by thiols, and none of the other transcription factors were activated by thiols. AP-1 activation was inhibited by catalase (500 U/ml) plus SOD plus ethanol (1 mM). Addition of 700 microM H2O2 also activated AP-1, and catalase at 500 U/ml prevented this activation. This indicates that H2O2 produced as a result of autooxidation of thiols can activate AP-1 but not NF-kappa B. Thus a close association between exposure to reducing agents, activation of NF-kappa B, and elevation of MnSOD gene expression is demonstrated.


Hypothalamic modulation of growth hormone secretion in the rhesus monkey: evidence from intracerebroventricular infusions of glucose, free fatty acid, and ketone bodies.
To evaluate the hypothalamus as a possible site of metabolic modulation of GH secretion, we studied the GH response to insulin hypoglycemia (IHG) and nicotinic acid (NA)-induced FFA depression in the absence and presence of third ventricular (ivt) infusions of glucose, oleic acid (Ol-Ac), or beta-hydroxybutyrate (beta OHB). Four rhesus monkeys had been prepared for chronic remote iv and ivt infusions as well as blood sampling from the adjacent room. Statistical evaluation used a two-way analysis of variance and individual comparisons with Tukey's Studentized range test. The GH response (area under the curve +/- SE) to IHG was significantly reduced by a concomitant ivt glucose infusion (control, 1.0 +/- 0.1; IHG, 12.1 +/- 3.3; IHG plus ivt glucose, 7.0 +/- 1.2 microgram/L.120 min). The GH response to FFA depression was significantly reduced by ivt Ol-Ac or beta OHB infusion (control, 6.0 +/- 1.0; NA, 51.5 +/- 4.1; Na plus Ol-Ac, 81.2 +/- 1.3; NA plus beta OHB, 38.6 +/- 3.5 microgram/L.300 min). Introcerebroventricular infusions of glucose, Ol-Ac, or beta OHB alone had no effect on plasma GH, glucose, FFA, or beta OHB concentrations. These results provide evidence for a hypothalamic site of metabolic modulation of GH secretion in the rhesus monkey. This does not exclude an additional effect directly at the pituitary gland.


Rigid Oxazole Acinetobactin Analog Blocks Siderophore Cycling in Acinetobacter baumannii.
The emergence of multidrug resistant (MDR) Gram-negative bacterial pathogens has raised global concern. Nontraditional therapeutic strategies, including antivirulence approaches, are gaining traction as a means of applying less selective pressure for resistance in vivo. Here, we show that rigidifying the structure of the siderophore preacinetobactin from MDR Acinetobacter baumannii via oxidation of the phenolate-oxazoline moiety to a phenolate-oxazole results in a potent inhibitor of siderophore transport and imparts a bacteriostatic effect at low micromolar concentrations under infection-like conditions.


Estrogen receptors in mast cells from arterial walls.
We examined the presence of estrogen receptors (ER) in vascular mast cells and a possible genomic effect of estrogens on the expression of mast cell (MC) mediators such as chymase, TNF alpha, NOS and IL-10, which are known to affect the course of atherosclerosis. Immunocytochemical detection of mast cell tryptase and the co-localization of ERs in MCs from abdominal aortic vessels from 10 fertile woman, 10 postmenopausal women and 15 men was performed. The genomic expression of IL-10, TNF alpha, and NOS was analyzed by RT-PCR and chymase activity by spectrophotometry after 24 h incubation with 17-beta estradiol (0.2-0.5 ng/mL) in rat purified peritoneal MCs. A similar number of MCs were found in both intima and adventitia layers from men, and fertile and postmenopausal women, while ERs were detected only in the arterial walls from fertile women. The mRNA expressions of IL-10 and TNF alpha, as well as chymase activity, were not affected. A moderate increment of NO and both NOS, and a reduction in TNF alpha cytotoxicity was observed after incubating peritoneal MCs with 17-beta estradiol at a concentration of 0.5 ng/mL. Taken together, these results indicate that vascular MCs express ERs. The data demonstrate that estrogens can directly modify vascular MC activity. This is a novel mechanism of synergistic cooperation for the protective role of estrogens in the genesis of atherosclerosis.


Photonic nanojets generated using square-profile microsteps.
We have shown experimentally that square-profile microsteps on a silica substrate, with square sides of 0.4, 0.5, 0.6, and 0.8 μm and height of 500 nm, illuminated through the substrate by a linearly polarized laser beam of wavelength λ=633 nm, produce, near the surface, enhanced-intensity regions (termed photonic nanojects), with their intensity being six times higher than that of the incident light and their respective full width at half-maximum diameters being 0.44λ, 0.43λ, 0.39λ, and 0.47λ, which is below the diffraction limit of 0.51λ. It is worth noting that when the step side is smaller than the wavelength, the focus is found within the step; otherwise the focus is outside the step, which is similar to an optical candle.


Relationship between ornithine decarboxylase and cytoskeletal organization in cultured human keratinocytes: cellular responses to phorbol esters, cytochalasins, and alpha-difluoromethylornithine.
Changes in cell shape occur during the cell cycle and influence cell proliferation and differentiation. In order to study how altered cell proliferation and cell shape are interrelated, we have studied ornithine decarboxylase (ODC) regulation in cultured normal human epidermal keratinocytes (NHEK). Cytoskeletal disruptors have been reported to modulate regulation of ODC; the products of ODC, the polyamines, influence actin polymerization rates in vitro, and polyamine auxotrophs have profoundly disrupted cytoskeletons. Therefore, altered ODC levels could be involved in signaling changes in cell shape and an intact cytoskeleton could transduce signals to regulate ODC levels. We had previously observed that the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), which profoundly alters cell shape, markedly suppresses ODC biosynthesis in NHEK solely at posttranscriptional/protein synthesis levels. TPA treatment caused NHEK to rapidly assume a rounded morphology that was accompanied by a change in actin organization, as determined by rhodamine-phalloidin labeling. Immunolocalization of ODC showed a perinuclear/nuclear distribution in untreated NHEK and a more diffuse pattern after TPA treatment that was apparent within 15-30 min. Changes in ODC enzyme activity are not significant until 60 min after TPA treatment. NHEK treated with cytochalasin B or D to inhibit actin polymerization exhibited a diffuse ODC localization that could be reversed by removal of the cytochalasin; inhibition of ODC by alpha-difluoromethylornithine caused a diffuse ODC localization. All treatments resulted in cytoskeletal remodeling. These data are the first evidence for a distinct subcellular localization for ODC and suggest that changes in ODC localization may be an initial step in regulation of ODC activity. Furthermore, changes in ODC activity cause an altered cytoskeleton, suggesting one means by which growth regulatory signals can be transduced to the cytoskeleton from various signaling pathways.


Lesions within the head direction system reduce retrosplenial c-fos expression but do not impair performance on a radial-arm maze task.
The lateral mammillary nuclei are a central structure within the head direction system yet there is still relatively little known about how these nuclei contribute to spatial performance. In the present study, rats with selective neurotoxic lesions of the lateral mammillary nuclei were tested on a working memory task in a radial-arm maze. This task requires animals to distinguish between eight radially-oriented arms and remember which arms they have entered within a session. Even though it might have been predicted that this task would heavily tax the head direction system, the lesion rats performed equivalently to their surgical controls on this task; no deficit emerged even when the task was made more difficult by rotating the maze mid-way through testing in order to reduce reliance on intramaze cues. Rats were subsequently tested in the dark to increase the use of internally generated direction cues but the lesion rats remained unimpaired. In contrast, the lateral mammillary nuclei lesions were found to decrease retrosplenial c-Fos levels. These results would suggest that the head direction system is not required for the acquisition of the standard radial-arm maze task. It would also suggest that small decreases in retrosplenial c-Fos are not sufficient to produce behavioural impairments.


Critical pedagogy in a health service management development programme: can "critically thinking" managers change the NHS management culture?
Management development programmes available to NHS managers focus on a performance orientation and sustain a culture of managerial and medical domination. This paper aims to question whether it is possible to consider NHS management development from a critical (empowerment culture) perspective. Features of the critical management studies approach (CMS) are identified. A new MSc is evaluated against these characteristics, examining the teaching and learning processes and students' perceptions of the programme. The aim is to develop critical thinkers who can return to their organizations and challenge existing power structures and practices to change local cultures and enhance health services. Empirical research employed anonymous student questionnaires and a focus group. Student evaluations suggest the MSc can deliver a critical pedagogy and help managers understand issues of power and empowerment, challenge dominant cultures, innovate and effect small, local changes in the NHS culture. There is a need to continue evaluating the programme and include other stakeholders. Longitudinal research should assess the impact of the managers' changed values, attitudes and behaviours on colleagues, clients and the local cultures. The paper identifies some of the tensions of developing "critical" health service managers, and the problems they encounter back in the "uncritical" NHS context, as well as some of the challenges in "facilitating" a critical curriculum. It questions the ethics of developing (or not) a critical perspective in a local context unfamiliar with CMS. Management development in the NHS largely ignores critical pedagogy. This paper makes a small and unique contribution to understanding how developing "critically thinking" managers can challenge the dominant culture. However, the limitations of such a small-scale study and ethical implications are noted.


Niche-Mediated Integrin Signaling Supports Steady-State Hematopoiesis in the Spleen.
Outside-in integrin signaling regulates cell fate decisions in a variety of cell types, including hematopoietic stem cells (HSCs). Our earlier published studies showed that interruption of periostin (POSTN) and integrin-αv (ITGAV) interaction induces faster proliferation in HSCs with developmental stage-dependent functional effects. In this study, we examined the role of POSTN-ITGAV axis in lymphohematopoietic activity in spleen that hosts a rare population of HSCs, the functional regulation of which is not clearly known. Vav-iCre-mediated deletion of Itgav in the hematopoietic system led to higher proliferation rates, resulting in increased frequency of primitive HSCs in the adult spleen. However, in vitro CFU-C assays demonstrated a poorer differentiation potential following Itgav deletion. This also led to a decrease in the white pulp area with a significant decline in the B cell numbers. Systemic deletion of its ligand, POSTN, phenocopied the effects noted in Vav-Itgav-/- mice. Histological examination of Postn-deficient spleen also showed an increase in the spleen trabecular areas. Importantly, these are the myofibroblasts of the trabecular and capsular areas that expressed high levels of POSTN within the spleen tissue. In addition, vascular smooth muscle cells also expressed POSTN. Through CFU-S12 assays, we showed that hematopoietic support potential of stroma in Postn-deficient splenic hematopoietic niche was defective. Overall, we demonstrate that POSTN-ITGAV interaction plays an important role in spleen lymphohematopoiesis.


A Biosensor-Based Leaf Punch Assay for Glutamine Correlates to Symbiotic Nitrogen Fixation Measurements in Legumes to Permit Rapid Screening of Rhizobia Inoculants under Controlled Conditions.
Legumes are protein sources for billions of humans and livestock. These traits are enabled by symbiotic nitrogen fixation (SNF), whereby root nodule-inhabiting rhizobia bacteria convert atmospheric nitrogen (N) into usable N. Unfortunately, SNF rates in legume crops suffer from undiagnosed incompatible/suboptimal interactions between crop varieties and rhizobia strains. There are opportunities to test much large numbers of rhizobia strains if cost/labor-effective diagnostic tests become available which may especially benefit researchers in developing countries. Inside root nodules, fixed N from rhizobia is assimilated into amino acids including glutamine (Gln) for export to shoots as the major fraction (amide-exporting legumes) or as the minor fraction (ureide-exporting legumes). Here, we have developed a new leaf punch based technique to screen rhizobia inoculants for SNF activity following inoculation of both amide exporting and ureide exporting legumes. The assay is based on measuring Gln output using the GlnLux biosensor, which consists of Escherichia coli cells auxotrophic for Gln and expressing a constitutive lux operon. Subsistence farmer varieties of an amide exporter (lentil) and two ureide exporters (cowpea and soybean) were inoculated with different strains of rhizobia under controlled conditions, then extracts of single leaf punches were incubated with GlnLux cells, and light-output was measured using a 96-well luminometer. In the absence of external N and under controlled conditions, the results from the leaf punch assay correlated with 15N-based measurements, shoot N percentage, and shoot total fixed N in all three crops. The technology is rapid, inexpensive, high-throughput, requires minimum technical expertise and very little tissue, and hence is relatively non-destructive. We compared and contrasted the benefits and limitations of this novel diagnostic assay to methods.


The C-terminal third of human respiratory syncytial virus attachment (G) protein is partially resistant to protease digestion and is glycosylated in a cell-type-specific manner.
The soluble form of the human respiratory syncytial virus (HRSV) attachment protein (Gs) was purified from the supernatant of infected cell cultures by immunoaffinity chromatography. Digestion of Gs with proteases and Western blot analysis identified two fragments that were partially resistant to protease degradation. Reactivity with diagnostic monoclonal antibodies located these two fragments in the primary structure of the G molecule. The large fragment spanned the C-terminal third of the G protein whereas the small fragment represented the N-terminal half of the large fragment. Purification of Gs from infected cells (either HEp-2 or M6) followed by protease digestion located host-cell-dependent glycosylation of the G protein in the unique part of the large protease-resistant fragment. The use of HRSV mutants encoding truncated G proteins allowed us to place some of the host-cell-dependent glycosylation differences in a small segment of the G protein. Interestingly, cell-specific glycosylations in the C-terminal half of the large protease-resistant fragment influenced the expression of certain epitopes located in its N-terminal half. These results bear important implications for the three-dimensional structure of the G glycoprotein.


The effects of endurance exercise combined with high-temperature head-out water immersion on serum concentration of brain-derived neurotrophic factor in healthy young men.
To evaluate acute changes in serum brain-derived neurotrophic factor (BDNF) concentration following combined endurance exercise and heat stress through head-out water immersion (HOI). Observational study with crossover design. Ten healthy young male participants performed HOI at 40 °C (40 °C HOI) or continuous cycling at 60% of maximal oxygen uptake while immersed in 40 °C (40 °C HOI-ex) or 23 °C water (23 °C HOI-ex) for 15 min. Serum BDNF, cortisol and lactate concentrations, and core temperature (Tcore) were measured pre, immediately post, and 15 and 30 min post-immersion. BDNF concentration increased immediately and 15 min after 40 °C HOI-ex, but not after 40 °C or 23 °C HOI-ex. No changes in Tcore concentration were observed during 23 °C HOI-ex (Pre; 37.3 °C ± 0.3 °C, Post; 37.8 °C ± 0.2 °C, Post 15; 37.4 °C ± 0.3 °C, Post 30; 37.2 °C ± 0.2 °C). Tcore increased significantly post, post 15, and post 30 min of 40 °C HOI (Pre; 37.1 °C ± 0.4 °C, Post; 38.8 °C ± 0.5 °C, Post 15; 37.9 °C ± 0.4 °C, Post 30; 37.9 °C ± 0.2 °C) and 40 °C HOI-ex (Pre; 37.2 °C ± 0.2 °C, Post; 40.2 °C ± 0.7 °C, Post 15; 38.9 °C ± 0.5 °C, Post 30; 38.3 °C ± 0.5 °C). Tcore was higher in 40 °C HOI-ex compared with 40 °C HOI and 23 °C HOI-ex immediately post and post 15 min. Plasma lactate and cortisol were significantly higher in 40 °C HOI-ex compared with 40 °C HOI and 23 °C HOI-ex after immersion (p = 0.001). While 15 min HOI alone or thermoneutral exercise do not increase BDNF concentration, both combined may form a time-efficient strategy to acutely elevate BDNF.


Localization of HSP47 mRNA in murine bleomycin-induced pulmonary fibrosis.
Heat shock protein 47 (HSP47) is a collagen-specific molecular chaperone that has been shown to play a major role in the processing and/or secretion of procollagen. However, the knowledge on which cells are actually synthesizing HSP47 in the lung parenchyma in pulmonary fibrosis was only limited. The aim of the present study was to investigate the localization of HSP47 messenger ribonucleic acid (mRNA) in normal lung and in the lungs of mice in bleomycin-induced pulmonary fibrosis, using in situ hybridization. For the purpose, ICR mice were intravenously injected with 10 mg/kg per day of bleomycin for five consecutive days. The lung cells expressing HSP47 mRNA were identified in control (saline alone) and bleomycin-treated mice by in situ hybridization. The signal for HSP47 mRNA was markedly increased in bleomycin-treated lungs compared with that of controls. HSP47 mRNA was localized in alpha-smooth-muscle-actin-positive myofibroblasts, surfactant-protein-A-positive type II pneumocytes, and F4/80-positive macrophages in the active fibrotic areas. These results suggest that these cells may synthesize procollagen in the fibrotic process of bleomycin-treated lungs through upregulation of HSP47 mRNA and play an important role in fibrogenesis.


The myth of core stability.
The principle of core stability has gained wide acceptance in training for the prevention of injury and as a treatment modality for rehabilitation of various musculoskeletal conditions in particular of the lower back. There has been surprisingly little criticism of this approach up to date. This article re-examines the original findings and the principles of core stability/spinal stabilisation approaches and how well they fare within the wider knowledge of motor control, prevention of injury and rehabilitation of neuromuscular and musculoskeletal systems following injury.


Distribution of KIR genes in the Iranian population.
Killer-cell immunoglobulin-like receptors (KIR) are a family of inhibitory and activating receptors that are expressed mainly by natural killer cells. The KIR gene family is highly polymorphic, and its genomic diversity is achieved through differences in gene content as well as allelic polymorphism. The number of KIR loci has been reported to be various among individuals and therefore resulting in different KIR haplotypes. This study represents the first report on the distribution of 17 presently defined KIR genes and pseudogenes in the Iranian population. In our study, 200 unrelated healthy individuals were KIR typed by a novel polymerase chain reaction-sequence-specific primers genotyping assay, and Iranian KIR genes distribution was compared with other ethnic groups. Over all, twenty-six different genotype profiles were found in our population and all KIR genes were observed. The most frequent non-framework KIR genes detected in our population were KIR2DL1 (96.5%), KIR3DL1 (91.5%), KIR2DS4 (91.5%) and the pseudogene KIR2DP1 (96.5%). The most commonly observed KIR genotype in Iranian population with a frequency of 27.5% consisted of KIR2DL1, KIR2DL3, KIR2DL4, KIR3DL1, KIR3DL2, KIR3DL3 and KIR2DS4 genes and the pseudogenes KIR2DP1 and KIR3DP1, which was compatible with a homozygote group-A haplotype. In addition, we found a new genotype (KIR2DL2, KIR2DL4, KIR2DL5, KIR3DL2, KIR3DL3, KIR2DS2, KIR2DS3, KIR2DS5, KIR3DS1 and KIR3DP1) in our samples. The results show that distribution of KIR genes in the Iranian population has common general features with the Caucasian populations studied before but still with unique, decreased or increased frequencies of several loci.


How do plants sense their nitrogen status?
The primary processes that contribute to the efficient capture of soil nitrate are the development of a root system that effectively explores the soil and the expression of high-affinity nitrate uptake systems in those roots. Both these processes are highly regulated to take into account the availability and distribution of external nitrate pools and the endogenous N status of the plant. While significant progress has been made in elucidating the early steps in sensing and responding to external nitrate, there is much less clarity about how the plant monitors its N status. This review specifically addresses the questions of what N compounds are sensed and in which part of the plant, as well as the identity of the signalling pathways responsible for their detection. Candidates that are considered for the role of N sensory systems include the target of rapamycin (TOR) signalling pathway, the general control non-derepressible 2 (GCN2) pathway, the plastidic PII-dependent pathway, and the family of glutamate-like receptors (GLRs). However, despite significant recent progress in elucidating the function and mode of action of these signalling systems, there is still much uncertainty about the extent to which they contribute to the process by which plants monitor their N status. The possibility is discussed that the large GLR family of Ca2+ channels, which are gated by a wide range of different amino acids and expressed throughout the plant, could act as amino acid sensors upstream of a Ca2+-regulated signalling pathway, such as the TOR pathway, to regulate the plant's response to changes in N status.


[Unstable angina pectoris. Pathogenesis, treatment and prognosis].
Unstable angina pectoris is a term used for various clinical conditions of angina pectoris, such as new onset of angina, accelerated angina, resting angina and post-infarction angina. The natural history varies according to clinical presentation, and the prognosis differs considerably for the different types of unstable angina. The first step in the pathophysiological mechanisms leading to unstable angina is thought to be rupture of an atherosclerotic plaque. Exposure of the content of the plaque leads to varying degrees of coronary obstruction, due to coronary thrombosis and vasoconstriction. A key element is dysfunction and activation of the endothelium. Patients with unstable angina should be hospitalized and treated with anti-anginal drugs, and with aspirin and heparin to reduce risk of myocardial infarction. Coronary angiography should be considered to identify patients who might benefit from angioplasty or coronary bypass surgery. Reduction of risk factors is important for long-term outcome.


Implementing a multiplex-PCR test for the diagnosis of acute gastroenteritis in hospitalized children: Are all enteric viruses the same?
Multiplex-PCR is a valuable tool for diagnosing viral acute gastroenteritis (AGE), enabling the detection of multiple pathogens. However, distinguishing between active disease and shedding poses challenges. This study aimed to evaluate viral AGE epidemiology and compare clinical characteristics among the five most common viruses. Rotavirus vaccine was introduced in 2010, with 70% coverage achieved in southern Israel in two years. All rectal swabs for multiplex-PCR targeting rotavirus, norovirus, adenovirus, astrovirus and sapovirus from hospitalized diarrheic children <5 years were included, from December 2017 through March 2022. Detection of the same virus within two months was considered a single episode. Clinical analysis included episodes with single-virus detection and negative bacterial PCR. Among 5,879 rectal swabs, 2,662 (45.3%) tested positive for at least one virus, with 245 (9.2%) showing multiple virus detection. Rotavirus was the most prevalent. While rotavirus exhibited typical winter-spring seasonality in 2018-19, an unusual off-season surge was observed during the second year of the COVID-19 pandemic. Among negative bacterial PCR episodes, 34.6% had mucus stool, 5.9% had bloody stool, and 29.3% received antibiotics. Astrovirus or sapovirus infections were associated with higher rates of hospital-acquired AGE and immunodeficiency (P<0.05), whereas rotavirus infections had higher rates of dehydration severity and acute kidney injury (P<0.05). Enteric viruses were detected in 45.3% of rectal swabs from hospitalized children with diarrhea. Despite vaccination efforts, rotavirus remained prevalent and caused more severe disease. Continuous surveillance using multiplex-PCR is crucial for accurate management and future prevention strategies for viral AGE.


A New Model for Predicting Hypothyroidism After Intensity-Modulated Radiotherapy for Nasopharyngeal Carcinoma.
To develop a model that can predict the risk of hypothyroidism (HT) after intensity-modulated radiotherapy (IMRT) for nasopharyngeal carcinoma (NPC), and to accordingly recommend dose constraints. NPC patients treated between 2011 and 2015 were retrospectively reviewed. HT was defined by an abnormally high level of thyrotropin. The dosimetry parameters Vx (percentage of thyroid volume receiving more than x Gy of radiation) and Va,b (percentage of thyroid volume receiving >a Gy, while ≤b Gy radiation) were calculated. The primary endpoint was the development of HT within the first 2 years after IMRT. The least absolute shrinkage and selection operator and multivariate logistic regression were used to identify predictors of HT. A total of 545 patients were included in the analyses, with a median follow-up of 36 months. Of the 545 patients, 138 developed HT within 2 years, and the 2-year incidence of HT was 25.3%. In patients with thyroid volume >20 cm3, the 2-year incidence of HT was 11.7% (16/137); in patients with thyroid volume ≤20 cm3 and V30,60 ≤ 80%, the 2-year HT incidence was 19.9% (33/166); in patients with thyroid volume ≤20 cm3 and V30,60 > 80%, the 2-year incidence of HT was 36.8% (89/242). Thyroid volume and V30,60 could be reliable predictors of HT after IMRT for NPC. For patients with thyroid volume ≤20 cm3, thyroid V30,60 ≤ 80% might be a useful dose constraint to adopt during IMRT planning.


Effect of depression on psychomotor skills, eye movements and recognition-memory.
In this study 12 depressed outpatients were compared to 12 healthy controls with respect to their performance on a number of cognitive tasks, including a recognition-memory task, and their eye movements and pupil size were recorded while watching a traffic film. The recognition-memory task consisted of words with intermediate hedonic tone (neutral words), words with high hedonic tone ('good' words) and words with low hedonic tone ('bad' words). Patients performed slower on perceptual-motor tasks which could be characterized as effort-demanding, while no difference between groups was found on effortless tasks. In addition, the range of horizontal eye movements, an indication of visual span, was found to be less in patients. Signal-detection analysis on the recognition-memory data showed an impairment of 'pure' memory in depressives. Analyses on response bias indicated that patients had more false alarms, but only with respect to good words. It is concluded that patients exhibit cognitive deficits, including memory impairment, a narrower visual span and a risky response strategy on good words, which may be an indication of the trouble patients have in processing emotionally toned words.


Lymphocyte proliferative response to allergen in timothy sensitive patients and healthy persons.
Presented are the results of the evaluation of lymphocytes proliferative response to a specific allergen in 30 pollinosis patients with hypersensitivity to timothy pollen, confirmed by the subject's allergy history, diagnostic skin tests, allergen-specific histamine release from basophils and by the presence of allergen-specific IgE-antibodies. The level of spontaneous proliferation of lymphocytes in pollinosis patients was found to increase. Manifestations of cell responses to Con A very similar in diseased and healthy subjects. The allergen-induced proliferative response of lymphocytes in pollinosis patients did not differ essentially from that in healthy persons. The authors make the supposition that the application of LBTR (using 3H-Thymidine incorporation) in demonstrating the immediate hypersensitivity to pollen allergens does not prove to be well grounded.


Renewable Butanol Production via Catalytic Routes.
Fluctuating crude oil price and global environmental problems such as global warming and climate change lead to growing demand for the production of renewable chemicals as petrochemical substitutes. Butanol is a nonpolar alcohol that is used in a large variety of consumer products and as an important industrial intermediate. Thus, the production of butanol from renewable resources (e.g., biomass and organic waste) has gained a great deal of attention from researchers. Although typical renewable butanol is produced via a fermentative route (i.e., acetone-butanol-ethanol (ABE) fermentation of biomass-derived sugars), the fermentative butanol production has disadvantages such as a low yield of butanol and the formation of byproducts, such as acetone and ethanol. To avoid the drawbacks, the production of renewable butanol via non-fermentative catalytic routes has been recently proposed. This review is aimed at providing an overview on three different emerging and promising catalytic routes from biomass/organic waste-derived chemicals to butanol. The first route involves the conversion of ethanol into butanol over metal and oxide catalysts. Volatile fatty acid can be a raw chemical for the production of butanol using porous materials and metal catalysts. In addition, biomass-derived syngas can be transformed to butanol on non-noble metal catalysts promoted by alkali metals. The prospect of catalytic renewable butanol production is also discussed.


Growth hormone, insulin-like growth factor 1, and insulin signaling-a pharmacological target in body wasting and cachexia.
Cachexia is an irreversible process that can develop in the course of chronic disease. It is characterized by the remodeling of the metabolic, inflammatory, and endocrine pathways. Insulin, growth hormone (GH), and insulin-like growth factor 1 (IGF-1) are involved in glucose, protein, and fat metabolism, which regulates body composition. In body wasting and cachexia, their signaling is impaired and causes anabolic/catabolic imbalance. Important mechanisms include inflammatory cytokines and neurohormonal activation. Remodeled post-receptor insulin, GH, and IGF-1 pathways constitute a potential target for pharmacological treatment in the setting of body wasting and cachexia. Peroxisome proliferator-activated receptor gamma agonists, drugs inhibiting angiotensin II action (angiotensin II antagonists and inhibitors of angiotensin-converting enzyme), and testosterone, which interfere with post-receptor pathways of insulin, GH, and IGF-1, were investigated as pharmacological intervention targets and various clinically important implications were reported. There are several other potential targets, but their treatment feasibility and applicability is yet to be established.


Crohn's disease presenting as vulvar edema in a 15-year-old girl.
Vulval involvement is an uncommon extraintestinal manifestation of Crohn's disease, and it is very rare in children. Patients with vulval CD typically present with erythema and edema of the labia majora, which can progresses to extensive ulcer formation. Vulval CD can appear before or after intestinal problems or it may occur simultaneously. We present a 15-years-old girl with bilateral labial hypertrophy which revealed a Crohn's disease. The course of her lesion was independent of the intestinal disease and responded significantly to medical treatment including Mesalamine, corticosteroid and local care. We emphasize that although vulval involvement in childhood is uncommon, Crohn's disease must be considered in the differential diagnosis of nontender, red, edematous lesions of the genital area.


Microstructure Changes in Polyester Polyurethane upon Thermal and Humid Aging.
The microstructure of compression molded Estane 5703 films exposed to 11%, 45%, and 80% relative humidity and 70 °C for 1 and 2 months has been studied by small-angle neutron scattering (SANS), Fourier transform infrared spectroscopy (FTIR), gel permeation chromatography (GPC), and differential scanning calorimetry (DSC). Scattering data indicated increase of the interdomain distance and domain size with a higher humidity and longer aging time. GPC data showed a progressive shortening of polyurethane chains with increasing humidity and aging time. The shortening of the polyurethane chains caused a drop of the glass transition temperature of soft segments, and promoted crystallization of the soft segments during long-time storage of the aged samples at room temperature. FTIR showed a substantial increase in the number of inter-urethane H-bonds in the aged samples. This correlates with the increase of the hard domain size and the degree of phase separation as measured by SANS. The data collected reveals that the reduced steric hindrance caused by hydrolysis of ester links in polybutylene adipate residues promotes the organization of hard segments into domains, leading to the increase of domain size and distance, as well as phase segregation in aged Estane. These findings provide insight into the effects of humidity and thermal aging on the microstructure of aged polyester urethane from molecular to nanoscale level.


[Individuals and changes in health organizations: a psychosociological approach].
The Brazilian health sector has undergone a severe crisis, affecting the case-resolving capacity, efficiency and governability of the health system as a whole and health organizations in particular. Although innovative management systems and tools have been encouraged, such innovations are limited in their ability to spawn organizational change, especially with regard to the challenge of enabling individual adherence to institutional projects and relations involving individuals and organizations. This paper focuses on the French psychosociological approach for analyzing and intervening in organizations, one of whose main thinkers is Eugène Enriquez. In its view of contemporary organizations, this approach focuses on the conflict between reproduction and creation as the main problem to be solved by management processes. While an organization is essentially seen as a place of order and repetition, organizational change implies the challenge of bringing creative individuals into the organization's project, avoiding the trap of controlling their minds and behavior.


Auditory and facial nerve dysfunction in patients with hemifacial microsomia.
Hemifacial microsomia (HFM) is a common craniofacial disorder characterized by a wide spectrum of anomalies, including conductive hearing loss due to external and middle ear deformities. However, the prevalence of sensorineural hearing loss (SNHL) as well as facial nerve dysfunction is underappreciated. To determine the frequency of auditory and facial nerve dysfunction and its relationship to more severe forms of bilateral HFM. Retrospective medical record review to characterize the clinical severity of HFM and the prevalence and nature of the associated auditory and facial nerve dysfunction. Center for Craniofacial Anomalies at the University of California, San Francisco, Medical Center. Ninety-nine pediatric patients with HFM evaluated at the University of California, San Francisco, Medical Center. The prevalence of SNHL and facial nerve dysfunction in this patient population and any associations between these 2 characteristics. Hearing loss was present in 74 (75%) of 99 patients, with a conductive component in 73 patients. Sensorineural hearing loss was present in 11 patients ( 11%), with mixed hearing loss in most patients. Fourteen patients required rehabilitation with auditory amplification. Nearly a quarter of the patients (22 [22%] of 99) had facial nerve dysfunction, but only 1 patient had facial palsy on the same side as the SNHL. There was a statistically significant association between having auricular abnormalities and conductive hearing loss or SNHL (P = .30 and .80, respectively). However, there was no statistically significant association between bilateral HFM and the occurrence of either SNHL or facial paralysis, nor was there an association between auditory and facial nerve dysfunction. Sensorineural hearing loss and facial nerve dysfunction are common in HFM. These findings have important implications in the treatment of patients with HFM.


Induction of resistance to 2',2'-difluorodeoxycytidine in the human ovarian cancer cell line A2780.
2',2'-Difluorodeoxycytidine (gemcitabine; dFdC) is a nucleoside analog with promising antitumor activity. To be active it must be phosphorylated by deoxycytidine kinase (dCK). We induced resistance to gemcitabine in the human ovarian carcinoma cell line A2780 by exposure to increasing concentrations of gemcitabine. At 72 hours' exposure the IC50, defined as the concentration of gemcitabine causing 50% growth inhibition, increased from 0.6 nmol/L gemcitabine in A2780 to 92 mumol/L in the resistant variant, AG6000. AG6000 is cross-resistant to other drugs that require activation by dCK, such as I-beta-D-arabinofuranosylcytosine, 5-aza-2'-deoxycytidine, and 2-chlorodeoxyadenosine. AG6000 was also cross-resistant to 2',2'-difluorodeoxyuridine (dFdU), the deamination product of gemcitabine. In addition, cross-resistance to the multidrug-resistance drugs doxorubicin and vincristine was observed. This was not associated with induction of P-glycoprotein. No accumulation of gemcitabine triphosphate could be detected in AG6000 cells, in contrast to the parental A2780 cells. There was no specific dCK activity in extracts from AG6000 cells. Western blot analysis using a polyclonal anti-dCK antibody did not reveal any dCK protein in AG6000 cell extracts. Reverse transcribed and polymerase chain reaction amplified mRNA, using specific dCK primers, demonstrated that AG6000 expressed a normal length amplicon of 701 base pairs, besides an aberrant amplicon of 500 base pairs. Although the resistant cell line is routinely cultured in 6 mumol/L gemcitabine, the resistant phenotype can be maintained for at least 10 passages without gemcitabine. These results indicate that the gemcitabine resistance phenotype is stable and mainly due to dCK deficiency.


Clinical characteristics and treatment outcomes in patients with Helicobacter pylori-positive chronic immune thrombocytopenic purpura.
Immune thrombocytopenic purpura (ITP) is the condition caused by increased platelet destruction and or decreased platelet production. Previous studies have demonstrated the association and efficacy of Helicobacter pylori (H. pylori) eradication therapy in patients with chronic ITP. Data in Thai patients, however, are limited. A prospective cross-sectional analytic study was conducted in adult patients with chronic ITP to determine the prevalence and clinical predictive factors of H. pylori infection and evaluate the efficacy of H. pylori eradication therapy. H. pylori-infected patients received eradication therapy (omeprazole 40 mg/day, clarithromycin 1000 mg/day, amoxicillin 2000 mg/day) for 2 weeks. The platelet counts at baseline and monthly for 6 months after the end of treatment were evaluated. Of the 25 patients, 9 patients (36%) had H.pylori infection. H. pylori infection is higher among women than men. There were two clinical factors included 1) relapsed ITP 2) response after the first-line treatment statistically proven to be associated with H. pylori infection with an odds ratio and p value of 7.7, p = 0.035 and ND (not determined due to small sample size), p < 0.001. Nearly 80% of infected patients had the platelet count response after eradication therapy with the median time to response of 4 months. The prevalence of H. pylori infection is modest in Thai adult patients with chronic ITP. A history of relapsed ITP and high quality of response after first-line treatment indicated H. pylori infection. Therefore, the urea breath test should be recommended in patients who have a relapsed ITP condition with a history of good response after first-line therapy.


A dynamic phase transition model for spatial agglomeration processes.
A nonlinear model of population migration is presented in order to provide a dynamic explanation for the formation of metropolitan areas. "In Section 2 the model is introduced in terms of the rate equations for the mean values of the regional population numbers with specifically chosen individual transition rates. Section 3 gives a survey of concepts and results for the convenience of the reader not interested in the details of the mathematical derivations. Section 4 derives the stationary solutions of the rate equations, that is, the equilibria of the system. Section 5 treats the time dependent solutions of the model equations focussing on the exact analytic solutions along so-called symmetry paths. Section 6 analyzes the dynamic stability of the symmetry path solutions and decides which stationary states are unstable and which are stable equilibrium states."


Analyzing Health-Related Quality of Life Data to Estimate Parameters for Cost-Effectiveness Models: An Example Using Longitudinal EQ-5D Data from the SHIFT Randomized Controlled Trial.
The aim of this article is to discuss methods used to analyze health-related quality of life (HRQoL) data from randomized controlled trials (RCTs) for decision analytic models. The analysis presented in this paper was used to provide HRQoL data for the ivabradine health technology assessment (HTA) submission in chronic heart failure. We have used a large, longitudinal EuroQol five-dimension questionnaire (EQ-5D) dataset from the Systolic Heart Failure Treatment with the I f Inhibitor Ivabradine Trial (SHIFT) (clinicaltrials.gov: NCT02441218) to illustrate issues and methods. HRQoL weights (utility values) were estimated from a mixed regression model developed using SHIFT EQ-5D data (n = 5313 patients). The regression model was used to predict HRQoL outcomes according to treatment, patient characteristics, and key clinical outcomes for patients with a heart rate ≥75 bpm. Ivabradine was associated with an HRQoL weight gain of 0.01. HRQoL weights differed according to New York Heart Association (NYHA) class (NYHA I-IV, no hospitalization: standard care 0.82-0.46; ivabradine 0.84-0.47). A reduction in HRQoL weight was associated with hospitalizations within 30 days of an HRQoL assessment visit, with this reduction varying by NYHA class [-0.07 (NYHA I) to -0.21 (NYHA IV)]. The mixed model explained variation in EQ-5D data according to key clinical outcomes and patient characteristics, providing essential information for long-term predictions of patient HRQoL in the cost-effectiveness model. This model was also used to estimate the loss in HRQoL associated with hospitalizations. In SHIFT many hospitalizations did not occur close to EQ-5D visits; hence, any temporary changes in HRQoL associated with such events would not be captured fully in observed RCT evidence, but could be predicted in our cost-effectiveness analysis using the mixed model. Given the large reduction in hospitalizations associated with ivabradine this was an important feature of the analysis. The Servier Research Group.


Bis(N-methylindolyl)methane-based chemical probes for Hg2+ and Cu2+ and molecular IMPLICATION gate operating in fluorescence mode.
Bis(N-methylindolyl)methane derivatives behave as chemosensors for Hg(2+) and Cu(2+) ions relying on the absorption as well as emission changes. Rarely reported IMPLICATION logic gate operating in the fluorescence mode has been developed with respect to the emission band of one of the derivatives as output, with Hg(2+) and I(-) as inputs.


Approaches to AIDS education for the grassroots in Nigeria.
The most important aspect of reaching people at the grassroots level is to ensure their understanding by using the most appropriate language, and by ensuring that the largely illiterate population will, nevertheless, be well served by print and electronic mass media. The article also emphasizes the need to adapt educational aids to the audience they address, as well as the preference of offering interactive education.


Utilization of a Mobile Dental Vehicle for Oral Healthcare in Rural Areas.
Oral diseases remain one of the major global public health challenges, and the worldwide urban⁻rural disparities in oral health are significant. Residents in rural areas generally suffer from a higher prevalence and severity of dental caries and periodontal disease, yet they face numerous difficulties and barriers in accessing oral healthcare. Conventional strategies, such as building of dental clinics or, hospitals, or the provision of outreach services by using disposable materials, are neither practical nor effective in rural settings. Mobile dental vehicles (MDVs) have been proposed as an alternative strategy to supplement the traditional oral healthcare in many regions. They have usually been utilized in school-based oral health programs, providing dental care to the homeless or migrants, and screening programs for the population for various oral diseases. Due to their high mobility, MDVs are particularly valuable for the underserved populations living in rural areas. The advance of dental devices enables MDVs to be operated in a self-sufficient manner. This allows the MDV to function almost as well as a conventional dental clinic, providing a variety of dental treatments, including scaling, restoration, and oral surgery. This article discusses the use of MDVs as a solution to urban⁻rural inequality in receiving oral healthcare.


Effect of hydration on the stability of the collagen-like triple-helical structure of [4(R)-hydroxyprolyl-4(R)-hydroxyprolylglycine]10.
X-ray analysis has been carried out on a crystal of the collagen model peptide (Hyp(R)-Hyp(R)-Gly)10 [where Hyp(R) is 4(R)-hydroxyproline] with 1.5 A resolution. The triple-helical structure of (Hyp(R)-Hyp(R)-Gly)10 has the same helical parameters and Rich and Crick II hydrogen bond patterns as those of other collagen model peptides. However, our full-length crystal structure revealed that almost all consecutive Hyp(R) residues take the up-up pucker in contrast to putative down-up puckering propensities of other collagen model peptides. The unique feature of thermodynamic parameters associated with the conformational transition of this peptide from triple helix to single coil is that both enthalpy and entropy changes of the transition are much smaller than those of other model peptides such as (Pro-Pro-Gly)10 and (Pro-Hyp(R)-Gly)10. To corroborate the precise structural information including main- and side-chain dihedral angles and intra- and interwater bridge networks, we estimated the degrees of hydration by comparing molecular volumes observed experimentally in solution to those calculated ones from the crystal structure. The results showed that the degree of hydration of (Hyp(R)-Hyp(R)-Gly)10 is comparable to that of (Pro-Hyp(R)-Gly)10 in the triple-helical state, but the former was more highly hydrated than (Pro-Hyp(R)-Gly)10 in the single-coil state. Because hydration reduces the enthalpy due to the formation of a hydrogen bond with a water molecule and diminishes the entropy due to the restriction of water molecules surrounding a peptide molecule, we concluded that the high thermal stability of (Hyp(R)-Hyp(R)-Gly)10 is able to be described by its high hydration in the single-coil state.


Theoretical and experimental study of structural slow light in a microfiber coil resonator.
In this paper, a compact slow-light microfiber coil resonator (MCR) is fabricated and the slow-light properties of it are analyzed and tested. Based on coupled-wave theory, a theoretical model for describing the slow-light propagation in the MCR is established. Experimentally, the MCR slow-light element is fabricated and its relative slow-light time delay is measured. The group velocity of the light pulse in the MCR slow-light element can be reduced to about 0.47c (c is the speed of light in vacuum) and the shape of the light pulse passing through the MCR is well preserved.


Guanosine primes acute myeloid leukemia for differentiation via guanine nucleotide salvage synthesis.
Differentiation arrest represents a distinct hallmark of acute myeloid leukemia (AML). Identification of differentiation-induction agents that are effective across various subtypes remains an unmet challenge. GTP biosynthesis is elevated in several types of cancers, considered to support uncontrolled tumor growth. Here we report that GTP overload by supplementation of guanosine, the nucleoside precursor of GTP, poises AML cells for differentiation and growth inhibition. Transcriptome profiling of guanosine-treated AML cells reveals a myeloid differentiation pattern. Importantly, the treatment compromises leukemia progression in AML xenograft models. Mechanistically, GTP overproduction requires sequential metabolic conversions executed by the purine salvage biosynthesis pathway including the involvement of purine nucleoside phosphorylase (PNP) and hypoxanthine phosphoribosyltransferase 1 (HPRT1). Taken together, our study offers novel metabolic insights tethering GTP homeostasis to myeloid differentiation and provides an experimental basis for further clinical investigations of guanosine or guanine nucleotides in the treatment of AML patients.


Tolerance of gastric mucosal flap to postoperative irradiation.
When malignant lesions of the oral cavity, base of tongue, and oropharynx are treated with radical resection, adequate reconstruction is required. The free gastric mucosal flap with microvascular transfer is being used with increasing frequency at Washington University Medical Center. Because of the advanced nature of the primary lesions, most patients also require postoperative radiation therapy. In this paper the tolerance of the gastric mucosal flap to postoperative radiation therapy is reviewed. The changes resulting from radiation therapy in the mucosal flap were found to be acceptable, and no major complications were encountered.


[The multimedia workstation, electronic assistant to the health care professional].
The health professional multimedia workstation is the natural entry point to health and knowledge networks. It should allow an easy and transparent management of all the data necessary for patient management. Multimedia nature of processed information reflects evolution of medical techniques that involve more and more complex objects such as video sequences or digitized signals. Workstations can be considered from 3 points of view: the professional end-users, the developers and the decision makers. The long term goal for the end-user is to establish an easy access in a working environment, that gives him/her the feeling of a single comprehensive application running on a single computer when the information system relies on a set of heterogeneous and geographically distributed components. Development of a workstation requires the integration into the same environment of tools to localize, access, manipulate and communicate the required information within a health information network. Analysis of the 445 Medline-indexed publications for the January 1991 to December 1994 period, that included the word workstation either in their title or in their abstract, helps in refining objectives and challenges both for the health professionals and the decision makers.


Bacterial adherence to tantalum versus commonly used orthopedic metallic implant materials.
Evaluation of bacterial adhesion to pure tantalum and tantalum-coated stainless steel versus commercially pure titanium, titanium alloy (Ti-6Al-4V), and grit-blasted and polished stainless steel. Experimental in vitro cell culture study using Staphylococcus aureus and Staphylococcus epidermidis to evaluate qualitatively and quantitatively bacterial adherence to metallic implants. A bacterial adhesion assay was performed by culturing S. aureus (ATCC 6538) and S. epidermidis (clinical isolate) for one hour with tantalum, tantalum-coated stainless steel, titanium, titanium alloy, grit-blasted and polished stainless steel metallic implant discs. Adhered living and dead bacteria were stained using a 2-color fluorescence assay. Adherence was then quantitatively evaluated by fluorescence microscopy and digital image processing. Qualitative adherence of the bacteria was analyzed with a scanning electron microscope. The quantitative data were related to the implant surface roughness (Pa-value) as measured by confocal laser scanning microscopy. Bacterial adherence of S. aureus varied significantly (p = 0.0035) with the type of metallic implant. Pure tantalum presented with significantly (p < 0.05) lower S. aureus adhesion compared to titanium alloy, polished stainless steel, and tantalum-coated stainless steel. Furthermore, pure tantalum had a lower, though not significantly, adhesion than commercially pure titanium and grit-blasted stainless steel. Additionally, there was a significantly higher S. aureus adherence to titanium alloy than to commercially pure titanium (p = 0.014). S. epidermidis adherence was not significantly different among the tested materials. There was no statistically significant correlation between bacterial adherence and surface roughness of the tested implants. Pure tantalum presents with a lower or similar S. aureus and S. epidermidis adhesion when compared with commonly used materials in orthopedic implants. Because bacterial adhesion is an important predisposing factor in the development of clinical implant infection, tantalum may offer benefits as an adjunct or alternative material compared with current materials commonly used for orthopedic implants.


In vitro inhibition of human MMP-2 collagenolytic and gelatinolytic activity by neutralizing monoclonal antibodies.
Matrix metalloproteinases (MMPs) have been implicated in the degradation of the extracellular matrix in normal and pathological tissue remodelling. Among the MMPs, MMP-2 is the most commonly studied protease that has been involved in cancer, inflammation, infective diseases, degenerative diseases of the brain and vascular diseases. In this study, monoclonal antibodies (MAbs) were generated against human MMP-2, purified, characterized and tested for their ability to inhibit the enzymatic activity of MMP-2. Out of 12 positive clones generated against MMP-2, 2 clones (F2-1-11 and G8-25-5) were selected for further characterization. The selected clones react specifically with human pro and active form of MMP-2 in enzyme linked immunosorbant assay (ELISA), dot immunobinding assay (DIA) and Western blot and do not cross react with other human metalloproteinases or MMP-2 from other species. Additionally, these MAbs (F2-1-11 and G8-25-5) selectively inhibit collagenolytic and gelatinolytic activity of APMA ((p-aminophenylmercuric acetate)-activated-pro-MMP-2 and MMP-2, respectively.


Identification, characterization and crystal structure analysis of the human spliceosomal U5 snRNP-specific 15 kD protein.
The U5 small ribonucleoprotein particle (snRNP) contains various proteins involved in catalytic activities mediating conformational rearrangements of the spliceosome. We have isolated and characterized the evolutionarily highly conserved human U5 snRNP-specific protein U5-15kD. The crystal structure of U5-15kD determined at 1.4 A resolution revealed a thioredoxin-like fold and represents the first structure of a U5 snRNP-specific protein known so far. With respect to human thioredoxin the U5-15kD protein contains 37 additional residues causing structural changes which most likely form putative binding sites for other spliceosomal proteins or RNA. Moreover, a novel intramolecular disulfide bond replaces the canonical one found in the thioredoxin family. Even though U5-15kD appears to lack protein disulfide isomerase activity, it is strictly required for pre-mRNA splicing in vivo as we demonstrate by genetic depletion of its ortholog in Saccharomyces cerevisiae. Our data suggest that the previously reported involvement of its Schizosaccharomyces pombe ortholog Dim1p in cell cycle regulation is a consequence of its essential role in pre-mRNA splicing.


Imaging of the optic chiasm and retrochiasmal visual pathways.
The exploration of the chiasmal and retrochiasmal visual pathways is based on magnetic resonance imaging. A bitemporal hemianopsis suggests a lesion of the optic chiasm while homonymous lateral hemianopsis should lead to a search for a lesion of the retrochiasmal visual pathways. The causes of chiasmal impairment are mainly tumoral. The exploration protocol is based on MRI with T1-weighted sagittal sections, then T2- and T1-weighted coronal sections with and without injection. In case of a retrochiasmal syndrome, the MRI exploration protocol is a function of the type of occurrence of the deficiency and the context.


Health-promoting and disease-mitigating potential of Verbascum thapsus L. (common mullein): A review.
Medicinal plants represent rich sources of traditional medicines and numerous currently used medicines are either directly or indirectly derived from plants. Verbascum thapsus L. (great mullein or common mullein), a medicinal herb indigenous to northern Africa, western and central Asia, and Europe, has been brought to the Americas and Australia. It has been used as a medicine for lung, skin and throat disorders and has a long history of therapeutic importance, particularly as an astringent and calming agent. Presently, the dried leaves, flowers, various plant extracts and flower oil are used in several formulations within Indian traditional medicine. An extract taken from the roots is useful in minimizing toothache, and it also relieves stiffness and seizures. V. thapsus contains a wide variety of phytoconstituents, such as flavonoids, iridoid, phenylethanoid and phenylpropanoid glycosides, saponins, as well as vitamin C and minerals. The most valuable constituents are coumarin and hesperidin, which possess healing properties. Emerging literature based on experimental studies on V. thapsus demonstrates various biological and pharmacological properties, including antiviral, antioxidant, analgesic, sedative, anti-inflammatory, hypnotic, antibacterial, antifungal, as well as anticancer activities. The present review provides an updated, comprehensive, and critical evaluation of various health-promoting and disease-mitigating properties of V. thapsus.


Neuropathology of two Brazilian autopsied cases of tropical spastic paraparesis / HTLV-I associated myelopathy (TSP/HAM) of long evolution.
We report on a neuropathological analysis of two cases of TSP/HAM originating from Brazil. These two cases had, respectively, an evolution of 13 and 40 years. The main neuropathological findings consisted of spinal cord atrophy, mainly the lower thoracic cord, diffuse degeneration of the white and grey matter, rare foci of mononuclear and perivascular cuffs, and hyaline hardening of arteriolae. The supraspinal structures were normal, excepting for a slight gliosis in the cerebellum. An analysis on the long evolutive cases as described in the literature is outlined in this study.


Differential roles for three conserved histidine residues within the large subunit of carbamoyl phosphate synthetase.
Three conserved histidine residues, His-243, His-781, and His-788, located within the large subunit of carbamoyl phosphate synthetase from Escherichia coli were identified by sequence identity comparisons. These three histidine residues were individually mutated to asparagine residues. The H243N mutant enzyme was found to be critical for carbamoyl phosphate synthesis as the mutant protein was unable to synthesize carbamoyl phosphate at a significant rate (< 1/1500). By analysis of the effects of this mutation on the partial reactions catalyzed by CPS, it was determined that this mutation blocked the formation of the carbamate intermediate from carboxyphosphate and ammonia. The H781N mutant enzyme had an order of magnitude reduction for both the rate of carbamoyl phosphate formation and ATP synthesis which is consistent with the proposal that the carboxyl-terminal half of the large subunit is primarily involved in the phosphorylation of the putative carbamate intermediate. This mutation also reduced the effects of the allosteric activator ornithine on the Km parameters for ATP in the overall biosynthetic reaction and ADP in the ATP synthesis reaction. The H788N mutant enzyme is a functional protein which maintains the ability to synthesize carbamoyl phosphate at a rate comparable to that of the wild-type enzyme. The effects of this mutation are 10-fold reductions of the ATP synthetase and the bicarbonate-dependent ATPase activities with substantial increases in the Km values for ATP in the full biosynthetic reaction and for ADP in the ATP synthesis reaction.(ABSTRACT TRUNCATED AT 250 WORDS)


Heart failure in Poland: Left ventricular assist device destination therapy and other challenges of interventional cardiology and cardiac surgery.
Patients with severe heart failure (HF), who are not eligible for cardiac transplantation and receive optimal medical management, based mainly on the use of pharmacological treatment and devices such as resynchronization therapy (implantable cardioverter-defibrillator), achieve poor clinical outcomes and constitute a group with extremely poor prognosis. Currently, the technology used in the latest generation left ventricular assist devices (LVADs), such as the HeartMate 3, makes it possible to achieve patient survival at the level obtained by patients after heart transplantation, and they can be used not only in patients eligible for heart transplantation as a bridge to transplant, but also in those with significantly worse prognosis, who are ineligible for heart transplantation as destination therapy. The objective of this publication is to present recommendations from experts in cardiology and cardiac surgery, supported by clinical trial results, on the use of LVADs as a destination therapy in HF patients who are not eligible for cardiac transplantation. The paper also presents the issue of cardiac transplantation and extracorporeal membrane oxygenation therapy in Poland, as well as current challenges faced by interventional cardiology and cardiac surgery in Poland.


Detection of Aneuploidy in Cerebrospinal Fluid from Patients with Breast Cancer Can Improve Diagnosis of Leptomeningeal Metastases.
Detection of leptomeningeal metastasis is hampered by limited sensitivities of currently used techniques: MRI and cytology of cerebrospinal fluid (CSF). Detection of cell-free tumor DNA in CSF has been proposed as a tumor-specific candidate to detect leptomeningeal metastasis at an earlier stage. The aim of this study was to investigate mutation and aneuploidy status in CSF-derived cell-free DNA (cfDNA) of patients with breast cancer with a clinical suspicion of leptomeningeal metastasis. cfDNA was isolated from stored remnant CSF and analyzed by targeted next-generation sequencing (NGS; n = 30) and the modified fast aneuploidy screening test-sequencing system (mFAST-SeqS; n = 121). The latter method employs selective amplification of long interspaced nuclear elements sequences that are present throughout the genome and allow for fast and cheap detection of aneuploidy. We compared these results with the gold standard to diagnose leptomeningeal metastasis: cytology. Leptomeningeal metastasis was cytology proven in 13 of 121 patients. Low DNA yields resulted in insufficient molecular coverage of NGS for the majority of samples (success rate, 8/30). The mFAST-SeqS method, successful in 112 of 121 (93%) samples, detected genome-wide aneuploidy in 24 patients. Ten of these patients had cytology-proven leptomeningeal metastasis; 8 additional patients were either concurrently diagnosed with central nervous system metastases by radiological means or developed these soon after the lumbar puncture. The remaining six cases were suspected of leptomeningeal metastasis, but could not be confirmed by cytology or imaging. Aneuploidy was associated with development of leptomeningeal metastasis and significantly worse overall survival. Aneuploidy in CSF-derived cfDNA may provide a promising biomarker to improve timely detection of leptomeningeal metastasis.


Primary Ewing's sarcoma of the cranium.
We analyzed the data for a series of 14 patients with primary Ewing's sarcomas of the cranium who were treated since 1985. Our aim was to assess the long-term outcomes and the selection of appropriate treatment methods. The patients were reviewed with respect to their clinical presentations, treatment, and outcomes. Computed tomographic scanning of the brain was performed for all patients. Skeletal surveys with routine radiographs and technetium-99 bone scans to detect extracranial Ewing's sarcomas were performed for all patients. For all 14 patients, radical tumor excision was achieved surgically. All patients were then subjected to adjuvant multidrug chemotherapy and radiotherapy. The follow-up periods ranged from 8 months to 8 years (mean, 4.25 yr). The predominant presenting features were headaches, increased intracranial pressure, and scalp swelling. Excision was nearly total for nine patients and total for five patients. All patients experienced uneventful postoperative courses. One patient experienced a local recurrence, which was detected 2 years after surgery. This recurrent tumor was completely excised, and additional chemotherapy was administered. Eight of the 14 patients (57.1%) studied have survived 5 years or longer. Although primary Ewing's sarcoma of the cranium is a malignant bone tumor, it is associated with a good prognosis when treated with radical surgery, aggressive multidrug chemotherapy, and radiotherapy.


[New advances in the diagnosis of liver fibrosis].
Various chronic liver diseases may progress to liver fibrosis or liver cirrhosis. Assessment of the degree of liver fibrosis helps with clinical decision-making, prognosis prediction, and evaluation of therapeutic effect, and therefore, accurate diagnosis and evaluation of the degree of liver fibrosis is a research hotspot at present. The advances in the diagnosis of liver fibrosis in recent years include the publication of international consensuses and guidelines, evaluation of the outcome of liver fibrosis after antiviral therapy, development of the diagnosis of liver fibrosis in patients with non-viral hepatitis, and constant update of noninvasive diagnostic techniques.


[Hematological changes induced by erythrocytapheresis].
Although automated cell separators (apheresis) have undergone a lot of technical refinements, the effect of the procedure on hematological indices of donors is rarely taken into account. The purpose of this study is to identify potential hematologic changes in donors undergoing erythrocytapheresis. 30 apparently healthy adult donors were evaluated. Erythrocytapheresis procedure was performed using automated equipment. Hematologic measurements (hemoglobin, hematocrit, white blood cells counts and platelets) were analyzed before and after erythrocytapheresis in all donors. We observed a significant decrease in the donors in hemoglobin (p <0.0001), hematocrit (p <0.0001), leukocytes (p <0.0001), lymphocytes (p = 0.0267), and platelets (p <0.0001). On the other hand, we found no changes in segmented, monocytes, eosinophils and basophils post erythrocytapheresis. In this study we found a significant drop in complete blood count in blood donation procedure by erythrocytapheresis; there are hematological changes in both red and white cells in all donors; however, none of donors manifested symptoms of thrombocytopenia or anemia. This study demonstrates hematological changes post-donation and therefore requires larger multicenter studies, in order to establish guidelines for donors' safety in apheresis and also help in assessing donor suitability, especially given the present trend of double product apheresis collections.


Effect of adhesive layer thickness and drug loading on estradiol crystallization in a transdermal drug delivery system.
The effects of adhesive layer thickness and drug loading on estradiol crystallization were studied in a drug-in-adhesive patch. Patches containing different estradiol loadings (1.1% and 1.6% w/w) in different thicknesses (45, 60, and 90 μm) were prepared by coating of a homogenous mixture of adhesive solution and the drug on a siliconized release liner by a film applicator. After drying, the film was laminated on a Poly(ethylene terephthalate) backing layer and cut into appropriate size. Release tests were performed using thermostated Chien-type diffusion cells. Cross-section of the patches was observed by optical microscopy. Scanning electron microscopy was done for surface analysis of the patches after drug release test. Crystal formation was not expected in the adhesive layer based on the linear free-energy relationship formalisms however; crystalline regions were observed in different locations through the thickness of the patches. These regions were significantly more discontinuous in 45 μm samples which elucidated the effective role of adhesive layer thickness in drug crystallization. Extensive crystallization observed for thicker patches was attributed to the strong crosslinking capability of estradiol hemihydrate. Drug release study confirmed some of the crystallization results. No significant increase was observed in the burst release with increasing in thickness from 45 to 60 μm which can be attributed to the severe increase in the crystallization extent. Also, formation of a crystalline layer near the releasing surface and more discontinuous pattern of the crystals in some samples was confirmed by investigation of the drug release curves.


Evaluation of genome damage and its relation to oxidative stress induced by glyphosate in human lymphocytes in vitro.
In the present study we evaluated the genotoxic and oxidative potential of glyphosate on human lymphocytes at concentrations likely to be encountered in residential and occupational exposure. Testing was done with and without metabolic activation (S9). Ferric-reducing ability of plasma (FRAP), thiobarbituric acid reactive substances (TBARS) and the hOGG1 modified comet assay were used to measure glyphosate's oxidative potential and its impact on DNA. Genotoxicity was evaluated by alkaline comet and analysis of micronuclei and other nuclear instabilities applying centromere probes. The alkaline comet assay showed significantly increased tail length (20.39 microm) and intensity (2.19%) for 580 microg/ml, and increased tail intensity (1.88%) at 92.8 microg/ml, compared to control values of 18.15 mum for tail length and 1.14% for tail intensity. With S9, tail length was significantly increased for all concentrations tested: 3.5, 92.8, and 580 microg/ml. Using the hOGG1 comet assay, a significant increase in tail intensity was observed at 2.91 microg/ml with S9 and 580 microg/ml without S9. Without S9, the frequency of micronuclei, nuclear buds and nucleoplasmic bridges slightly increased at concentrations 3.5 microg/ml and higher. The presence of S9 significantly elevated the frequency of nuclear instabilities only for 580 microg/ml. FRAP values slightly increased only at 580 microg/ml regardless of metabolic activation, while TBARS values increased significantly. Since for any of the assays applied, no clear dose-dependent effect was observed, it indicates that glyphosate in concentrations relevant to human exposure do not pose significant health risk.


The effect of temperature upon the permeation of polar and ionic solutes through human epidermal membrane.
The temperature dependence of in vitro permeation through human epidermal membrane (HEM) was determined for urea, mannitol, tetraethylammonium ion (TEA), and corticosterone. The effect of temperature upon HEM electrical resistance was also measured. The majority of the experiments involved measuring the permeability coefficients of a specific permeant at 27 degrees C and 39 degrees C for a given HEM sample, the electrical resistance was also measured at each temperature. Similar experiments were also conducted with a model synthetic porous membrane. The effect of temperature was quantitated as the ratio of the permeability at 39 degrees C to the permeability at 27 degrees C for each permeant. These ratios observed for HEM with urea, mannitol, and TEA as the permeants were 1.66 +/- 0.05, 1.76 +/- 0.14, and 1.71 +/- 0.11, respectively. The change in temperature was shown to have a similar effect upon the electrical conductance of the HEM samples. The observed ratio for corticosterone permeation was 4.5 +/- 0.4. The experimental ratios observed for the three polar/ionic permeants were shown to approach those obtained from the model porous membrane and differed greatly from the ratio observed for the more lipophilic corticosterone, indicating differences in the effective transport mechanism/pathway for these classes of permeants. The permeability of urea was also observed to be inversely proportional to the electrical resistance of the HEM samples; this relationship was shown to be independent of temperature over the temperature range studied. The temperature dependence data and the observed relationship between urea permeability and electrical resistance strongly support the existence of a porous permeation pathway through the HEM as an operative diffusional route for polar-ionic permeants.


Spatiotemporal characteristics of the collisionless rf sheath and the ion energy distributions arriving at rf-biased electrodes.
A self-consistent fluid model is proposed for describing collisionless radio-frequency (rf) sheaths driven by a sinusoidal current source. This model includes all time-dependent terms in ion fluid equations, which are commonly ignored in some analytical models and numerical simulations. Moreover, an equivalent circuit model is introduced to determine self-consistently the relationship between the instantaneous potential at a rf-biased electrode and the sheath thickness. The time-dependent voltage wave form, the sheath thickness and the ion flux at the electrode as well as the spatiotemporal variations of the potential, the electric field and the ion density inside the sheath are calculated for various rf powers and ratios of the rf frequency to the ion plasma frequency. The ion energy distributions ( IEDs) impinging on the rf-biased electrode are also calculated with the ion flux at the electrode. The numerical results show that the frequency ratio is a crucial parameter determining the spatiotemporal variations of the rf sheath and the shape of the IEDs.


The reliability of electric pulp test after concussion injury.
Traumatic injuries to teeth present a difficult diagnostic problem with respect to determining the vitality of the affected teeth. Conventionally, a latency period of 4 to 6 weeks is proposed as the amount of time required for the return of a positive electric pulp test (EPT) response; however, this has been based on empirical data. The purpose of this study was to 1) determine a time frame during which a return of a reliable response to EPT may be measured, and 2) correlate this time frame with pulpal vascular changes that occur after traumatic injuries. Four canines from each of nine ferrets were used for this experiment (18 traumatized and 18 control teeth). A reproducible concussion injury was devised by releasing a known weight at a constant height above the facial surface of the experimental teeth. The EPT response was confirmed by the use of an EMG muscle recorder. Animals were killed at 24, 48 and 72 h and 6, 12 and 28 days after the injury and the teeth analyzed histologically. The histological sections showed an absence of inflammatory cells in both groups; however, according to the NIH image system, an increase in the number of vessels was detected in the experimental group when compared to the controls. In a majority of the cases, a positive response to the EPT returned within 12 days post concussion injury and as early as 10 days. In addition, a significant vascular change was accompanied with the loss of the EPT response. This study provided a reliable method by which concussion injuries may be studied using an animal model.


Portal hypertension and myeloproliferative neoplasms: a relationship revealed.
Background/Objectives. Patients with myeloproliferative neoplasms have a well-established increased risk of thrombosis. Many trials report identification of an underlying myeloproliferative neoplasm by investigation of the patients developing portal hypertensive esophagus and/or fundus variceal hemorrhage in the absence of any known etiology. This trial was designed to investigate the association between myeloproliferative neoplasms and portal hypertension and to detect the frequency of portal hypertension development in this subset of patients. Methodology. Twenty-nine patients previously diagnosed with polycythemia vera, essential thrombocytopenia, and primary myelofibrosis, who were under followup at the hematology outpatient clinic of our hospital, were included in the trial. Results. In our trial, we detected portal hypertension in 13.8% of the patients (n = 4), as a finding that was similar to those obtained in other studies performed to date. Conclusions. Considering the fact that diagnosis of myeloproliferative neoplasms usually takes a long time, treatment should be started (while, on the other hand, assessing the investigational and therapeutical choices for the complications) right after the bone marrow biopsy or cytogenetic studies required for establishing the final diagnosis have been performed.