Sister chromatid exchange in Polish White improved goats (Capra hircus).
The study was aimed at evaluating the frequency of spontaneous sister chromatid exchange in Polish White Improved goats (Capra hircus). The mean number of SCEs/cell was 2.73 +/- 1.84. The effect of sex and age on SCE incidence was also investigated. No statistically significant differences in the number of SCEs/cell were observed between the males and females. On the other hand, age was found to significantly influence SCE frequency. A lower SCE frequency was observed in younger goats. A positive correlation between chromosome length and SCE number was identified. The longer the chromosome, the more exchanges occurred. The highest number of SCEs was observed in the interstitial region, the lowest in the distal area.


Insulin-receptor phosphotyrosyl-protein phosphatases.
Calmodulin-dependent protein phosphatase has been proposed to be an important phosphotyrosyl-protein phosphatase. The ability of the enzyme to attack autophosphorylated insulin receptor was examined and compared with the known ability of the enzyme to act on autophosphorylated epidermal-growth-factor (EGF) receptor. Purified calmodulin-dependent protein phosphatase was shown to catalyse the complete dephosphorylation of phosphotyrosyl-(insulin receptor). When compared at similar concentrations, 32P-labelled EGF receptor was dephosphorylated at greater than 3 times the rate of 32P-labelled insulin receptor; both dephosphorylations exhibited similar dependence on metal ions and calmodulin. Native phosphotyrosyl-protein phosphatases in cell extracts were also characterized. With rat liver, heart or brain, most (75%) of the native phosphatase activity against both 32P-labelled insulin and EGF receptors was recovered in the particulate fraction of the cell, with only 25% in the soluble fraction. This subcellular distribution contrasts with results of previous studies using artificial substrates, which found most of the phosphotyrosyl-protein phosphatase activity in the soluble fraction of the cell. Properties of particulate and soluble phosphatase activity against 32P-labelled insulin and EGF receptors are reported. The contribution of calmodulin-dependent protein phosphatase activity to phosphotyrosyl-protein phosphatase activity in cell fractions was determined by utilizing the unique metal-ion dependence of calmodulin-dependent protein phosphatase. Whereas Ni2+ (1 mM) markedly activated the calmodulin-dependent protein phosphatase, it was found to inhibit potently both particulate and soluble phosphotyrosyl-protein phosphatase activity. In fractions from rat liver, brain and heart, total phosphotyrosyl-protein phosphatase activity against both 32P-labelled receptors was inhibited by 99.5 +/- 6% (mean +/- S.E.M., 30 observations) by Ni2+. Results of Ni2+ inhibition studies were confirmed by other methods. It is concluded that in cell extracts phosphotyrosyl-protein phosphatases other than calmodulin-dependent protein phosphatase are the major phosphotyrosyl-(insulin receptor) and -(EGF receptor) phosphatases.


In vitro DNA Inversions Mediated by the PsrA Site-Specific Tyrosine Recombinase of Streptococcus pneumoniae.
Site-specific recombination is a DNA breaking and reconstructing process that plays important roles in various cellular pathways for both prokaryotes and eukaryotes. This process requires a site-specific recombinase and direct or inverted repeats. Some tyrosine site-specific recombinases catalyze DNA inversions and regulate subpopulation diversity and phase variation in many bacterial species. In Streptococcus pneumoniae, the PsrA tyrosine recombinase was shown to control DNA inversions in the three DNA methyltransferase hsdS genes of the type I restriction-modification cod locus. Such DNA inversions are mediated by three inverted repeats (IR1, IR2, and IR3). In this work, we purified an untagged form of the PsrA protein and studied its DNA-binding and catalytic features. Gel retardation assays showed that PsrA binds to linear and supercoiled DNAs, containing or not inverted repeats. Nevertheless, DNase I footprinting assays showed that, on linear DNAs, PsrA has a preference for sites that include an IR1 sequence (IR1.1 or IR1.2) and its boundary sequences. Furthermore, on supercoiled DNAs, PsrA was able to generate DNA inversions between specific inverted repeats (IR1, IR2, and IR3), which supports its ability to locate specific target sites. Unlike other site-specific recombinases, PsrA showed reliance on magnesium ions for efficient catalysis of IR1-mediated DNA inversions. We discuss that PsrA might find its specific binding sites on the bacterial genome by a mechanism that involves transitory non-specific interactions between protein and DNA.


Photonic switching in waveguides using spatial concepts inspired by EIT.
Optical waveguide switches based on spatial arrangements inspired by electromagnetic induced transparency (EIT) concepts are presented. Interferometric control of optical signal by an optical gate is accomplished in three configurations. The first is employing a direct spatial version of EIT, the second is exploiting space reciprocity to accomplish performance not achievable in the time domain EIT and finally a novel version of EIT, using tunneling, is transformed into the spatial domain. For all configurations - closed form analysis as well as actual device simulation are presented.


Barriers to recommended gynecologic care in an urban United States HIV clinic.
Despite an increased risk for cervical cytologic abnormalities, HIV-infected women frequently miss their gynecology appointments. We examined barriers to adherence with gynecologic care in an urban HIV clinic. We conducted a cross-sectional survey of 200 women receiving gynecologic services in an urban HIV clinic, followed by focus groups. Primary outcomes included (1) missed gynecology appointments and (2) receipt of a Pap smear in the previous year. Independent variables included sociodemographic characteristics, child care responsibilities, substance use, depressive symptoms, social support, interpersonal violence, CD4 count, and HIV-1 RNA. We conducted multivariable logistic regression to examine associations between independent variables and outcomes. We then held two focus groups designed to gather opinions on and increase our understanding of the key findings from the survey. Of 200 women, 69% missed at least one gynecology appointment, and 22% had no Pap smear in the past year. In logistic regression, moderate (odds ratio [OR] 3.1, 95% confidence interval [CI] 1.4-6.7) and severe (OR 3.1, 95% CI 1.3-7.5) depressive symptoms and past-month substance use (OR 2.3, 95% CI 1.0-5.3) were associated with missing an appointment in the prior year. An education level of less than high school (OR 0.3, 95% CI 0.1-0.6) compared with high school diploma or greater was associated with not having a Pap smear in the previous year. When analyses were limited to women with a cervix (n = 166), moderate (OR 2.5, 95% CI 1.1-5.7) and severe (OR 2.5, 95% CI 1.0-6.3) depressive systems remained significantly associated with missing a gynecology appointment in the previous year and age >50 (OR 0.3, 95% CI 0.1-0.9), an HIV-1 RNA > 50 (OR 0.4, 95% CI 0.2-0.9), and education level less than high school (OR 0.2, 95% CI 0.1-0.5) were associated with not having a Pap smear in the past 12 months. Qualitative analysis of the focus group data suggested that fear, inclement weather, and forgetting appointments may contribute to missed gynecology appointments. Gynecologic healthcare is underused among HIV-infected women. We found that depressive symptoms, substance use, fear of the gynecologic examination, and simply forgetting about the appointment may be barriers to gynecologic care. Interventions targeting these barriers may improve use of gynecologic care among this population.


Rhizobium japonicum nitrogenase Fe protein gene (nifH).
A 12.1-kilobase PstI fragment from Rhizobium japonicum, which contains homology to both the Klebsiella pneumoniae and the Rhizobium meliloti nifH genes, was cloned into vector pHE3 . The nifH -homologous region was localized on the restriction enzyme cleavage map by Southern blot hybridization experiments. DNA fragments overlapping the R. japonicum nifH gene were subcloned into plasmid vectors to allow the expression of this region in Escherichia coli minicells. The nifH gene product (the polypeptide of the nitrogenase Fe protein) was thus found to have a molecular weight of 33,000. The complete nucleotide sequence of the nifH gene was established, and the amino acid sequence of its gene product was deduced. The reading frame is 882 nucleotides long, corresponding to 294 amino acids which add up to a polypeptide with a molecular weight of 31,525. There was extensive sequence homology with nifH genes or gene products from other nitrogen-fixing bacteria. The transcription initiation site of the R. japonicum nifH gene was found to lie 153 nucleotides upstream from the coding region and was preceded by a characteristic promoter sequence. A potential terminator region was located 13 nucleotides downstream from the coding region.


Bone mass accrual in children.
Bone accrual during childhood and adolescence is critical for the attainment of peak bone mass and is a major contributing factor towards osteoporosis in later life. Bone mass accrual is influenced by nonmodifiable factors, such as genetics, sex, race, ethnicity, and puberty, as well as modifiable factors, such as physical activity and diet. Recent progress in bone imaging has allowed clinicians and researchers to better measure the morphology, density, and strength of the growing skeleton, thereby encompassing key characteristics of peak bone strength. In this review, the patterning of bone accrual and contributors to these changes will be described, as well as new techniques assessing bone mass and strength in pediatric research and clinical settings. This review discusses factors influencing peak bone mass attainment and techniques used to assess the human skeleton. The rate of bone accrual and the magnitude of peak bone mass attainment occurs in specific patterns varying by sex, race, ethnicity, longitudinal growth, and body composition. Physical activity, diet, and nutritional status impact these processes. There is a need for longitudinal studies utilizing novel imaging modalities to unveil factors involved in the attainment and maintenance of peak bone strength.


Microwave-assisted extraction (MAE)-acid hydrolysis of dithiocarbamates for trace analysis in tobacco and peaches.
A simple and rapid method is presented for the analysis of residues of ethylenebis(dithiocarbamate) (maneb, zineb, and mancozeb) and N,N-dimethyldithiocarbamate (thiram and ziram) fungicides in dry tobacco leaves and peaches. Residues are extracted and hydrolyzed to CS(2) in a single step by use of microwave energy in a closed-vessel system while the evolved CS(2) trapped in a layer of iso-octane overlaying the reaction mixture is taken for gas chromatographic-flame photometric analysis. This combined extraction-hydrolysis step is carried out in 10 and 15 min for sets of 12 samples of tobacco and peach matrices, respectively. Total sample preparation time for GC analysis is 40 min. The limits of detection (LOD) are 0.005 mg/kg for thiram and ziram on peaches and 0.1 mg/kg for maneb, zineb, and mancozeb on tobacco. The respective LOD and limit of quantification (LOQ) levels in CS(2) equivalents are 0.003 and 0.006 mg/kg on peaches and 0.04 and 0.2 mg/kg on tobacco, respectively. Recoveries in the 0.01-60 mg/kg fortification range are 80-100% with respective relative standard deviations <20%. The method was used for the analysis of >3000 commercial tobacco samples including also different marketed cigarette brands.


A context-specific role for retinoblastoma protein-dependent negative growth control in suppressing mammary tumorigenesis.
The ability to respond to anti-growth signals is critical to maintain tissue homeostasis and loss of this negative growth control safeguard is considered a hallmark of cancer. Negative growth regulation generally occurs during the G0/G1 phase of the cell cycle, yet the redundancy and complexity among components of this regulatory network has made it difficult to discern how negative growth cues protect cells from aberrant proliferation. The retinoblastoma protein (pRB) acts as the final barrier to prevent cells from entering into the cell cycle. By introducing subtle changes in the endogenous mouse Rb1 gene (Rb1(ΔL)), we have previously shown that interactions at the LXCXE binding cleft are necessary for the proper response to anti-growth signals such as DNA damage and TGF-β, with minimal effects on overall development. This disrupts the balance of pro- and anti-growth signals in mammary epithelium of Rb1(ΔL/ΔL) mice. Here we show that Rb1(ΔL/ΔL) mice are more prone to mammary tumors in the Wap-p53(R172H) transgenic background indicating that negative growth regulation is important for tumor suppression in these mice. In contrast, the same defect in anti-growth control has no impact on Neu-induced mammary tumorigenesis. Our work demonstrates that negative growth control by pRB acts as a crucial barrier against oncogenic transformation. Strikingly, our data also reveals that this tumor suppressive effect is context-dependent.


Basal low antioxidant capacity correlates with cognitive deficits in early onset psychosis. A 2-year follow-up study.
The objective of the study is to examine the association of baseline total antioxidant status (TAS) and glutathione (GSH) levels with short- and long-term cognitive functioning in patients with early onset first-episode psychosis, comparing affective and non-affective psychoses. We analysed 105 patients with an early onset-first episode psychosis (age 9-17 years) and 97 healthy controls. Blood samples were taken at admission for measurement of TAS and GSH, and cognitive performance was assessed at baseline and at 2years of follow-up. Regression analysis was used to assess the relationship between TAS/GSH levels at baseline and cognitive performance at both time points, controlling for confounders. Baseline TAS and GSH levels were significantly lower in patients than healthy controls. In patients, baseline TAS was positively associated with the global cognition score at baseline (p=0.048) and two years later (p=0.005), while TAS was not associated with cognitive functioning in healthy controls. Further, baseline TAS in patients was specifically associated with the memory domain at baseline and with the memory and attention domains two years later. Stratifying by affective and non-affective psychoses, significant associations were only found between TAS and cognition in the non-affective psychosis group. Baseline GSH levels were not associated with cognitive functioning at either time point in either group. The antioxidant defence capacity in early onset first-episode psychotic patients is directly correlated with global cognition at baseline and at 2years of follow-up, especially in non-affective psychosis.


Association between Skin Carotenoid Score Measured with Veggie Meter® and Adherence to the Mediterranean Diet among Adolescents from Southern Italy.
The Veggie Meter® (Longevity Link Corporation, Salt Lake City, UT, USA), is a new portable device for the non-invasive and rapid detection of skin carotenoid content, which represents an acceptable biomarker for the evaluation of fruit and vegetable (FV) intake. FVs are important components of a healthy diet, including the Mediterranean Diet (MD), which is a plant-based dietary pattern. Here, we evaluated the adherence to the MD via the administration of two online food questionnaires, and we measured the skin carotenoid content using the Veggie Meter® in a cohort of 498 healthy adolescents (233 males and 265 females) from Southern Italy. Using KIDMED and the MD Pyramid tests to assess the adherence to the MD, we found an average adherence (5.43 ± 2.57 and 7.20 ± 1.93, respectively) to the MD in our sample population. Moreover, we observed that the skin carotenoid score was 364.75 ± 98.29, which was within the normal range and inversely related to the BMI (r = -0.1461, p = 0.0011). Similar results were observed when the population was categorized by sex. Interestingly, we demonstrated, for the first time, a positive correlation between the carotenoid score and the adherence to the MD assessed using both the KIDMED and MD Pyramid tests in the total population (r = -0.2926, p < 0.0001 and r = -0.1882, p < 0.0001, respectively). The same direct correlation was found in adolescents according to their sex. Our findings highlight the potential of the Veggie Meter® as a feasible and promising tool for evaluating adherence to the MD and, ultimately, to promote healthy eating habits among adolescents.


Genotoxic effects of benzyl isothiocyanate, a natural chemopreventive agent.
Benzyl isothiocyanate (BITC) is contained in cruciferous plants which are part of the human diet. Numerous reports indicate that BITC prevents chemically induced cancer in laboratory animals and it has been postulated that BITC might also be chemoprotective in humans. On the other hand, evidence is accumulating that this compound is a potent genotoxin in mammalian cells by itself. To further elucidate the potential hazards of BITC, we investigated its genotoxic effects in different in vitro genotoxicity tests and in animal models. In in vitro experiments [differential DNA repair assay with Escherichia coli, micronucleus assay with human HepG2 cells and single cell gel electrophoresis (SCGE) assay with hepatocytes and gastrointestinal tract cells] pronounced dose-dependent genotoxic effects were found at low dose levels (</=5 microg/ml). In contrast, substantially weaker effects were obtained in in vivo experiments with laboratory rodents: in the differential DNA repair assay with E.coli cells, only moderate genotoxic effects were seen in indicator cells recovered from various organs of mice after treatment with high doses (between 90 and 270 mg/kg), while in SCGE assay with rats a change in the DNA migration pattern was seen at a dose level of 220 mg/kg body wt. These findings indicate that BITC is detoxified under in vivo test conditions. This assumption was supported by the results of in vitro experiments which showed that the genotoxic effects of BITC are markedly reduced by bovine serum albumin and human body fluids such as saliva and gastric juice. Additional experiments carried out on the mechanistic aspects of the genotoxicity of BITC showed that this compound causes formation of thiobarbituric acid-reactive substances in HepG2 cells and that its DNA damaging properties are diminished by alpha-tocopherol, vitamin C, sodium benzoate and beta-carotene, indicating the possible involvement of free radicals in the genotoxicity of BITC. The doses of BITC required to cause measurable DNA damage in laboratory rodents exceeded by far the dietary exposure levels of humans, but are similar to those which were required to inhibit chemically induced cancer in earlier animal experiments.


The Wnt signaling pathway and mitotic regulators in the initiation and evolution of mantle cell lymphoma: Gene expression analysis.
For an accurate understanding of mantle cell lymphoma (MCL), molecular behavior could be staged into two major events: lymphomagenesis with the t(11;14) translocation (initiation), and evolution into a more aggressive form (transformation). Unfortunately, it is still unknown which genes contribute to each event. In this study, we performed cDNA microarray experiments designed based on the concept that morphologically heterogeneous MCL samples would provide insights into the role of aberrant gene expression for both events. A total of 15 MCLs were collected from the files, which include a total of 237 MCL patients confirmed by histology as CCND1-positive. We posited four stepwise morphological grades for MCL: MCL in situ, MCL with classical form (cMCL), MCL with aggressive form (aMCL), and MCL with intermediate morphology between classical and aggressive forms at the same site (iMCL). To identify genes involved in initiation, we compared the tumor cells of MCL in situ (n=4) with normal mantle zone B lymphocytes (n=4), which were selected by laser microdissection (LMD). To identify genes contributing to transformation, we selected the overlapping genes differentially expressed between both cMCL (n=4) vs. aMCL (n=5) and classical vs. aggressive areas in iMCL (n=2) obtained by LMD. A significant number of genes (n=23, p=0.016) belonging to the Wnt signaling pathway were differentially expressed in initiation. This specific activation was confirmed by immuno-histochemistry, as MCL in situ had nuclear localization of phosphorylated-β-catenin with high levels of cytoplasmic Wnt3 staining. For transformation, identified 60 overlapping genes included a number of members of the p53 interaction network (CDC2, BIRC5 and FOXM1), which is known to mediate cell cycle progression during the G2/M transition. Thus, we observe that the Wnt signaling pathway may play an important role in initial lymphomagenesis in addition to t(11;14) translocations, and that specific mitotic regulators facilitate transformation into more aggressive forms.


Nanoparticle conversion chemistry: Kirkendall effect, galvanic exchange, and anion exchange.
Conversion chemistry is a rapidly maturing field, where chemical conversion of template nanoparticles (NPs) into new compositions is often accompanied by morphological changes, such as void formation. The principles and examples of three major classes of conversion chemical reactions are reviewed: the Kirkendall effect for metal NPs, galvanic exchange, and anion exchange, each of which can result in void formation in NPs. These reactions can be used to obtain complex structures that may not be attainable by other methods. During each kind of conversion chemical reaction, NPs undergo distinct chemical and morphological changes, and insights into the mechanisms of these reactions will allow for improved fine control and prediction of the structures of intermediates and products. Conversion of metal NPs into oxides, phosphides, sulphides, and selenides often occurs through the Kirkendall effect, where outward diffusion of metal atoms from the core is faster than inward diffusion of reactive species, resulting in void formation. In galvanic exchange reactions, metal NPs react with noble metal salts, where a redox reaction favours reduction and deposition of the noble metal (alloying) and oxidation and dissolution of the template metal (dealloying). In anion exchange reactions, addition of certain kinds of anions to solutions containing metal compound NPs drives anion exchange, which often results in significant morphological changes due to the large size of anions compared to cations. Conversion chemistry thus allows for the formation of NPs with complex compositions and structures, for which numerous applications are anticipated arising from their novel catalytic, electronic, optical, magnetic, and electrochemical properties.


A preliminary characterization of the cytosolic glutathione transferase proteome from Drosophila melanogaster.
The cytosolic GST (glutathione transferase) superfamily has been annotated in the Drosophila melanogaster genome database. Of 36 genes, four undergo alternative splicing to yield a total of 41 GST proteins. In the present study, we have obtained the 41 transcripts encoding proteins by RT (reverse transcription)-PCR using RNA template from Drosophila S2 cells, an embryonic cell line. This observation suggests that all of the annotated DmGSTs (D. melanogaster GSTs) in the proteome are expressed in the late embryonic stages of D. melanogaster. To avoid confusion in naming these numerous DmGSTs, we have designated them following the universal GST nomenclature as well as previous designations that fit within this classification. Furthermore, in the cell line, we identified an apparent processed pseudogene, gste8, in addition to two isoforms from the Delta class that have been published previously. Only approximately one-third of the expressed DmGSTs could be purified by conventional GSH affinity chromatography. The diverse kinetic properties as well as physiological substrate specificity of the DmGSTs are such that each individual enzyme displayed a unique character even compared with members from the same class.


An introductory survey of helminth control practices in south africa and anthelmintic resistance on Thoroughbred stud farms in the Western Cape Province.
Fifty-one per cent of 110 questionnaires, designed for obtaining information on helminth control practices and management on Thoroughbred stud farms in South Africa, were completed by farmers during 2000. The number of horses per farm included in the questionnaire survey ranged from 15 to 410. Foals, yearlings and adult horses were treated with anthelmintics at a mean of 7.3 +/- 3.0, 6.6 +/- 2.7 and 5.3 +/- 2.3 times per year, respectively. An average of 3.4 different drugs were used annually, with ivermectin being used bymost farmers during 1997-2000. On 43% of farms the weights of horses were estimated by weigh band and 45% of farmers estimated visually, while both were used on 7% of farms and scales on the remaining 5%. Doses were based on average group weight on 50% ofthe farms and on individual weights on 46%. Forty-three per cent of farmers performed faecal egg count reduction tests (FECRT). Most farmers rotated horses between pastures and treated new horses at introduction. Faecal removal was practiced on 61% of farms and less than 50% of farmers used alternate grazing with ruminants. Faecal egg count reduction tests were done on 283 horses, using oxibendazole, ivermectin and moxidectin on 10,9 and 5 farms, respectively, in the Western Cape Province during 2001. While the efficacy of oxibendazole was estimated by FECRT to range from 0-88% and moxidectin from 99-100%, ivermectin resulted in a 100% reduction in egg counts. Only cyathostome larvae were recovered from post-treatment faecal cultures.


Conjugation of Polymer-Coated Gold Nanoparticles with Antibodies-Synthesis and Characterization.
The synthesis of polymer-coated gold nanoparticles with high colloidal stability is described, together with appropriate characterization techniques concerning the colloidal properties of the nanoparticles. Antibodies against vascular endothelial growth factor (VEGF) are conjugated to the surface of the nanoparticles. Antibody attachment is probed by different techniques, giving a guideline about the characterization of such conjugates. The effect of the nanoparticles on human adenocarcinoma alveolar basal epithelial cells (A549) and human umbilical vein endothelial cells (HUVECs) is probed in terms of internalization and viability assays.


Suicide rates among working-age adults in South Korea before and after the 2008 economic crisis.
Multiple studies have shown that macroeconomic factors are associated with changes in suicide rates. We investigated how changes in economic conditions associated with the recent economic crisis in South Korea influenced suicide rates among working-age adults. Time-series analyses were performed to examine the temporal associations of national unemployment rates and sex-employment-specific suicide rates in South Korea from 2003 to 2011, with particular attention to the increases of suicides that occurred during the recessionary period that began in 2008. We also compared the relative risk of suicide among different occupations. National unemployment rates were positively associated with suicide rates among employed and unemployed men and women, with a 2-month to 3 month lagged period. Significant increases of suicide rates among working-age adults during the recession were detected in most of the subgroups stratified by age, sex and employment status. Forty-three per cent of the increase of suicides was derived from the employed population. Compared with workers in elementary occupations, the relative risk of suicide for mangers increased by threefold during the recessionary period. Among those who were employed, half of the increases in suicides occurred among clerks and workers involved in sales and services. Changes in macroeconomic conditions are tied to population-level suicide risks for employed and unemployed persons. However, these associations vary depending on sex, employment status and occupational roles. In advance of future economic crises, it is important to develop prevention initiatives intended to reach the diverse populations potentially exposed to the adverse effects of sudden economic disruptions.


[Lysosomotropic agents as regulators of the activity of lysosomal hydrolases].
Activities of lysosomal enzymes acid phosphatase, N-acetyl-beta-D-glucosaminidase, beta-galactosidase, acid lipase and cathepsins B and D were studied after accumulation of neutral red, acridine orange chloroquine and daunorubicin in lysosomes of fibroblasts of the LSM substrain. All the drugs studied proved to be inhibitors of these enzymes except of daunorubicin, which stimulated acid lipase activity. Lysosomotropic drugs are considered as possible regulators of the activity of lysosomes.


Feeding habits of Chaoboridae larvae in a tropical Brazilian reservoir.
The diet of the four larval instars of a Chaoboridae species of Lake Monte Alegre, was evaluated through the analysis of the crop content of individuals caught at night in the lake. Peridinium and zooflagellates were the major components of the diet of instars I and II, whereas microcrustaceans, especially the cladoceran Bosmina, were most important for late instars. When the preferential prey decreased, in the course of the year, the diet was more diversified, increasing the ingestion of Peridinium by instars III and IV. There is a relationship between the mouth diameter of the larvae and the maximum width, height, or diameter of the ingested organisms.


Against all odds: genocidal trauma is associated with longer life-expectancy of the survivors.
Does surviving genocidal experiences, like the Holocaust, lead to shorter life-expectancy? Such an effect is conceivable given that most survivors not only suffered psychosocial trauma but also malnutrition, restriction in hygienic and sanitary facilities, and lack of preventive medical and health services, with potentially damaging effects for later health and life-expectancy. We explored whether genocidal survivors have a higher risk to die younger than comparisons without such background. This is the first population-based retrospective cohort study of the Holocaust, based on the entire population of immigrants from Poland to Israel (N = 55,220), 4-20 years old when the World War II started (1939), immigrating to Israel either between 1945 and 1950 (Holocaust group) or before 1939 (comparison group; not exposed to the Holocaust). Hazard of death - a long-term outcome of surviving genocidal trauma - was derived from the population-wide official data base of the National Insurance Institute of Israel. Cox regression yielded a significant hazard ratio (HR = 0.935, CI (95%) = 0.910-0.960), suggesting that the risk of death was reduced by 6.5 months for Holocaust survivors compared to non-Holocaust comparisons. The lower hazard was most substantial in males who were aged 10-15 (HR = 0.900, CI (95%) = 0.842-0.962, i.e., reduced by 10 months) or 16-20 years at the onset of the Holocaust (HR = 0.820, CI (95%) = 0.782-0.859, i.e., reduced by18 months). We found that against all odds genocidal survivors were likely to live longer. We suggest two explanations: Differential mortality during the Holocaust and "Posttraumatic Growth" associated with protective factors in Holocaust survivors or in their environment after World War II.


Renoprotective effect of breviscapine through suppression of renal macrophage recruitment in streptozotocin-induced diabetic rats.
Experimental and clinical evidence has consistently demonstrated that renal macrophage infiltration is one of the most important events for the progression of diabetic nephropathy. Breviscapine is a flavonoid extracted from the Chinese herb Erigeron breviscapus. Previously, it was shown that treatment with breviscapine attenuated renal injury in the diabetic rats. The purpose of this study is to investigate whether the renoprotective effect of breviscapine is through suppression of renal macrophage recruitment in diabetic rats. Diabetes was induced bystreptozotocin injection, and breviscapine was administered orally at a dose of 20 mg/kg/day for 8 weeks. Control rats received vehicle or breviscapine with the same schedule. Breviscapine treatment markedly inhibited both an increase of albuminuria and glomeruli hypertrophy and tubulointerstitial injury without modifying mean arterial blood pressure and creatinine clearance. Levels of malondialdehyde and protein kinase C activities were markedly higher and antioxidant enzyme activities such as superoxide dismutase, catalase as well as glutathione peroxidase were significantly lower in the kidneys of diabetic rats than of the control group, breviscapine administration markedly remitted these changes. ED-1-positive cells and expression of intercellular adhesion molecule-1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1) in glomeruli and tubulointerstitium were all markedly elevated but were significantly reduced by breviscapine. Western blot analysis noted that the expression of transforming growth factor beta1 protein was increased 1.8-fold in the kidney in diabetic rats, breviscapine treatment could reduce increased expression of TGF-beta1 protein by 47%. This study describes a novel mechanism by which breviscapine confers a renoprotective effect.


Hypoglycaemia following treatment of hyperkalaemia with insulin and dextrose.
We describe four patients who developed symptomatic hypoglycaemia following treatment of hyperkalaemia with insulin and dextrose. Two patients had a delayed onset of hypoglycaemia, between 5 and 6 hours after treatment despite use of a 'soluble' type of insulin. A review of the literature revealed a variety of insulin and dextrose regimes but no research to assess the metabolic effects of such therapy in patients with renal failure. The possibility of significant hypoglycaemia following use of insulin and dextrose in the recommended dosages is rarely mentioned. The cases we describe demonstrate that there is no 'correct' dose of insulin and dextrose to suit every circumstance. Regular blood glucose estimations should be performed in all patients receiving such therapy for hyperkalaemia.


Visuomotor system uses target features unavailable to conscious awareness.
After lesions to primary visual cortex, patients lack conscious awareness of visual stimuli. Interestingly, however, some retain the ability to make accurate judgments about the visual world (i.e., so-called blindsight). Similarly, damage to inferior occipitotemporal regions of cortex (e.g., lateral occipital cortex) can result in an inability to perceive object properties while retaining the ability to act on them (i.e., visual form agnosia). In the present work, we demonstrate that the ability to interact with objects in the absence of conscious awareness is not isolated to those with restricted neuropathologic conditions. Specifically, neurologically intact individuals are able to program and execute goal-directed reaching movements to a target object without awareness of extrinsic target properties; they accurately tune the dynamics of their movement and modulate it online without conscious access to features of the goal object. Thus, the planning and execution of actions are not dependent on conscious awareness of the environment, suggesting that the phenomenon of blindsight (and agnosia) reflect normal conditions of the visual system.


SAPHO Syndrome: Current Developments and Approaches to Clinical Treatment.
SAPHO syndrome (synovitis, acne, pustulosis, hyperostosis, and osteitis) is a rare autoimmune disease which, due to its clinical presentation and symptoms, is often misdiagnosed and unrecognized. Its main features are prominent inflammatory cutaneous and articular manifestations. Treatments with immunosuppressive drugs have been used for the management of SAPHO with variable results. To date, the use of anti-TNF-α agents has proved to be an effective alternative to conventional treatment for unresponsive or refractory SAPHO cases. TNF-α is a pro-inflammatory cytokine and pivotal regulator of other cytokines, including IL-1 β, IL-6, and IL-8, involved in inflammation, acute-phase response induction, and chemotaxis. IL-1 inhibition strategies with anakinra have shown efficacy as first and second lines of treatment. In this review, we will describe the main characteristics of biological drugs currently used for SAPHO syndrome. We also describe some of the promising therapeutic effects of ustekinumab, an antibody against the p40 subunit of IL-12 and IL-23, after failure of multiple drugs including anti-TNF-α and anakinra. We discuss the use and impact of the new anti-IL-1 antagonists involved in the IL-17 blockade, in particular for the most difficult-to-treat SAPHO cases.


Ecdysone-dependent proteolysis of an apical surface glycoprotein may play a role in imaginal disc morphogenesis in Drosophila.
An apical surface glycoprotein, designated gp125 for its apparent molecular weight of 125,000, appears in Ca2(+)-free, ionic detergent extracts of imaginal discs of Drosophila melanogaster in response to the steroid hormone, 20-hydroxyecdysone (20-HE). Gp125 is not synthesized in response to 20-HE, but results from modification of an existing macromolecule. Treatment of discs or larval epidermis with serine protease (e.g., trypsin) results in hormone-independent production of gp125. Antiserum raised to electrophoretically purified gp125 recognizes, in addition to gp125, two closely related glycoproteins with higher apparent molecular weights, gp200 and gp180. This family of glycoproteins is localized at the apical surface of imaginal disc cells and of the epidermal epithelium in embryos, larvae and prepupae. Ca2+ affects both the solubility and the proteolytic products of this family of glycoproteins. We discuss the possibility that gp125 is generated through the action of a hormonally controlled serine protease in a process that is necessary for disc morphogenesis.


Stopping transformed cancer cell growth by rigidity sensing.
A common feature of cancer cells is the alteration of kinases and biochemical signalling pathways enabling transformed growth on soft matrices, whereas cytoskeletal protein alterations are thought to be a secondary issue. However, we report here that cancer cells from different tissues can be toggled between transformed and rigidity-dependent growth states by the absence or presence of mechanosensory modules, respectively. In various cancer lines from different tissues, cells had over tenfold fewer rigidity-sensing contractions compared with normal cells from the same tissues. Restoring normal levels of cytoskeletal proteins, including tropomyosins, restored rigidity sensing and rigidity-dependent growth. Further depletion of other rigidity sensor proteins, including myosin IIA, restored transformed growth and blocked sensing. In addition, restoration of rigidity sensing to cancer cells inhibited tumour formation and changed expression patterns. Thus, the depletion of rigidity-sensing modules through alterations in cytoskeletal protein levels enables cancer cell growth on soft surfaces, which is an enabling factor for cancer progression.


Effects of age and caloric restriction on mitochondrial protein oxidative damage in mice.
The hypothesis that life-span extension by caloric restriction (CR) is contingent upon the attenuation of macromolecular oxidative damage was tested in two different strains of mice: the C57BL/6, whose life span is extended by CR, and the DBA/2, in which CR has relatively minor or no impact on longevity. Mice were fed ad libitum (AL) or restricted to 40% lesser food, starting at 4 months of age. Protein damage was measured as protein-linked adducts of 4-hydroxy-2-nonenal (HNE) and malondialdehyde (MDA) in skeletal muscle mitochondria at 6 and 23 months of age. Protein-HNE and -MDA content increased with age in C57BL/6 mice and CR significantly attenuated these augmentations. Metalloprotease 1, NADP-dependent mitochondrial malic enzyme (isoform 2) and citrate synthase were identified by mass spectroscopy to contain HNE/MDA adducts. DBA/2 mice exhibited little effect of age or CR on protein HNE/MDA content in skeletal muscle mitochondria. In contrast, protein-HNE levels in liver mitochondria showed a significant increase with age in AL-fed mice of both strains, and CR caused significant attenuation of this damage. Overall, results indicated that the age-related increase in protein oxidative damage and its abatement by CR are genotype- and tissue-specific, and not a universal phenomenon.


Prevalence of asthma and allergic diseases in Croatian children is increasing: survey study.
To estimate the prevalence of asthma, allergic rhinitis, and atopic dermatitis among school children in the region of Primorsko-goranska County in Croatia, and compare the results with data from other countries. The study was conducted during the 2001-2002 school year, in complete adherence to the Phase One protocol of the International Study of Asthma and Allergies in Childhood (ISAAC). The target population comprised two age groups (6-7 and 13-14 years) in the region of Primorsko-Goranska County in Croatia. Data were collected using standardized ISAAC written questionnaire and asthma video questionnaire. There were 1,634 participating children in the 6-7 age group (response rate 80.3%) and 2,194 participating children in the 13-14 age group (response rate 89.8%). Estimated 12-month prevalence rates of symptoms were: wheezing 9.7% and 8.4%, allergic rhinitis symptoms 16.9% and 17.5%, allergic rhinoconjunctivitis symptoms 5.6% and 6.7%, and atopic dermatitis symptoms 5.4% and 3.4%, for younger and older age group, respectively. Results suggest an increase in the prevalence of atopic disease symptoms in north-west part of Croatia over the last few decades when compared to prior studies. The results are suitable for international comparison, suggesting that this part of Croatia is a county with a moderate prevalence of atopic diseases in the pediatric population. The results represent a baseline for further epidemiological research of asthma and allergic diseases.


Linkage rate between data from health checks and health insurance claims in the Japan National Database.
Japan's National Database (NDB) includes data on health checks and health insurance claims, is linkable using hash functions, and is available for research use. However, the linkage rate between health check and health insurance claims data has not been investigated. Linkage rate was evaluated by comparing observed medical and pharmaceutical charges among health check recipients in fiscal year (FY) 2009 (N = 21 588 883) with expected charges from the same population when record linkage was complete. Using the NDB, observed charges were estimated from the first published result of linking health check recipients in FY2009 and their health insurance claims in FY2010. Expected charges were estimated by combining 3 publicly available datasets, including data from the Medical Care Benefit Survey and an ad-hoc report by the Japan Health Insurance Association. Only 14.9% of expected charges were linked by the NDB. The linkage rate was higher for women than for men (18.2% vs 12.4%) and for elderly adults as compared with younger adults (>25% vs <10%). The linkage rate in the NDB was so low that any research linking health check and health insurance claims will not be reliable. Causes for the low linkage rate include differences between health check and health insurance claims data in name format (eg, insertion of a space between family and given names) and date of birth (Japanese vs Gregorian calendar). Investigation of the causes for the low linkage rate and measures for improvement are urgently needed.


Nerve growth factor-regulated emergence of functional delta-opioid receptors.
Sorting of intracellular G-protein-coupled receptors (GPCRs) either to lysosomes for degradation or to plasma membrane for surface insertion and functional expression is a key process regulating signaling strength of GPCRs across the plasma membrane in adult mammalian cells. However, little is known about the molecular mechanisms governing the dynamic process of receptor sorting to the plasma membrane for functional expression under normal and pathological conditions. In this study, we demonstrate that delta-opioid receptor (DOPr), a GPCR constitutively targeted to intracellular compartments, is driven to the surface membrane of central synaptic terminals and becomes functional by the neurotrophin nerve growth factor (NGF) in native brainstem neurons. The NGF-triggered DOPr translocation is predominantly mediated by the signaling pathway involving the tyrosine receptor kinase A, Ca(2+)-mobilizing phospholipase C, and Ca(2+)/calmodulin-dependent protein kinase II. Importantly, it requires interactions with the cytoplasmic sorting protein NHERF-1 (Na(+)/H(+) exchange regulatory factor-1) and N-ethyl-maleimide-sensitive factor-regulated exocytosis. In addition, this NGF-mediated mechanism is likely responsible for the emergence of functional DOPr induced by chronic opioids. Thus, NGF may function as a key molecular switch that redirects the sorting of intracellularly targeted DOPr to plasma membrane, resulting in new functional DOPr on central synapses under chronic opioid conditions.


Home health service utilization by Hispanic elderly immigrants: a longitudinal study.
Based on a longitudinal study of Hispanic elderly immigrants, factors associated with use of home health services (HHS) were examined. Interviews were conducted in a community study of 156 Hispanic elderly immigrants in 1983 and again in 1988 with 76 survivors using the Older Americans Resources and Services (OARS) questionnaire. Predisposing and illness level factors were significantly related to the use of HHS, however, enabling factors were not significant. A discriminant analysis indicated that a set of variables (gender, age, physical health, mental health, ADL function and Self Assessed Health) accounted for 54% of the variance. The strongest discriminators in use of home health services were age, ADL function and Self Assessed Health.


Bone cells predispose bone surfaces to resorption by exposure of mineral to osteoclastic contact.
The cell-free endocranial surface of young adult rat parietal bones was used as a substrate for osteoclastic bone resorption, either without prior treatment, or after incubation of the parietal bones with collagenase or neonatal rat calvarial cells. Untreated, the endocranial surface consisted of unmineralized organic fibres; incubation with calvarial cells or collagenase caused disruption and removal of these fibres, with extensive exposure of bone mineral on the endocranial surface, without morphologically detectable mineral dissolution. Neonatal rabbit osteoclasts resorbed bone to a greater extent from parietal bones pre-incubated with calvarial cells or collagenase than from untreated bones; mineral exposure and subsequent osteoclastic resorption were both increased if calvarial cells were incubated with parathyroid hormone; removal of bone mineral after incubation with calvarial cells removed the predisposition to osteoclastic resorption. These experiments demonstrate that calvarial cells are capable of osteoid destruction, and indicate that one mechanism by which osteoblasts induce osteoclastic bone resorption may be through digestion of the unmineralized organic material that covers bone surfaces, to expose the underlying resorption-stimulating bone mineral to osteoclastic contact.


[RAD29 and RAD31--new genes from Saccharomyces cerevisiae yeasts, participating in control of DNA repair. II. Clarification of possible functions of these genes].
Possible functions of previously described genes RAD29 and RAD31 involved in DNA repair were determined by analyzing the interaction between these genes and mutations in the genes of the three basic epistatic groups: RAD3 (nucleotide excision repair), RAD6 (error-prone mutagenic repair system), RAD52 (recombination repair pathway), and also the apn1 mutation that blocks the synthesis of major AP endonuclease (base excision repair). The results obtained in these studies and the estimation of the capability for excision repair of lesions induced by 8-metoxipsoralen and subsequent exposure to long-wavelength UV light in mutants for these genes led to the assumption that the RAD29 and RAD31 genes are involved in yeast DNA repair control.


Update on chemical peels.
The area of chemical peeling has seen an explosion of growth and interest in the last 5 years. The use of peeling for the therapy of pigmentary dyschromias, actinic keratoses and rhytides, depressed scarring, and in selected cases, irradiation dermatitis, is amplified by the discovery of new wounding agents and new variations in technique. Appropriate growth in the objective evaluation of these agents and techniques is dependent on the demonstrative histology and the sharing of knowledge of both excellent results and significant complications in the dermatologic surgical literature.


Stability of cetuximab and panitumumab in glass vials and polyvinyl chloride bags.
The stability of cetuximab and panitumumab in glass vials and polyvinyl chloride (PVC) bags stored at 4 degrees C for up to 14 days was studied. Sixty milliliters of cetuximab was drawn directly from a commercially available vial, and 20 mL was injected into each of three sterile, empty, 100-mL PVC bags through a preattached 0.22-microm filter. Three milliliters of panitumumab and 21 mL of 0.9% sodium chloride injection were mixed, and 8 mL of the mixture was injected into each of three empty PVC bags through a preattached 0.22-microm filter. Samples were analyzed immediately after preparation and again after storage at 4 degrees C for 7 and 14 days. Cetuximab and panitumumab concentrations were measured using a modification of a previously described enzyme-linked immunosorbent assay method. Stability was defined as the mean concentrations of the test solutions being within assay variability of the initial concentration. Solution appearance and color were assessed by observing the samples against black and white backgrounds. The percentages of initial concentration of cetuximab and panitumumab were over 90% stored at 4 degrees C after 14 days. No changes in color or turbidity were observed in any of the vials and the prepared solutions. Cetuximab 2 mg/mL was stable when stored for 14 days in a glass vial and in PVC bags at 4 degrees C. Panitumumab 20 mg/mL in a glass vial and 2.5 mg/mL in 0.9% sodium chloride injection in PVC bags was also stable when stored for 14 days at 4 degrees C.


HNF4A is essential for specification of hepatic progenitors from human pluripotent stem cells.
The availability of pluripotent stem cells offers the possibility of using such cells to model hepatic disease and development. With this in mind, we previously established a protocol that facilitates the differentiation of both human embryonic stem cells and induced pluripotent stem cells into cells that share many characteristics with hepatocytes. The use of highly defined culture conditions and the avoidance of feeder cells or embryoid bodies allowed synchronous and reproducible differentiation to occur. The differentiation towards a hepatocyte-like fate appeared to recapitulate many of the developmental stages normally associated with the formation of hepatocytes in vivo. In the current study, we addressed the feasibility of using human pluripotent stem cells to probe the molecular mechanisms underlying human hepatocyte differentiation. We demonstrate (1) that human embryonic stem cells express a number of mRNAs that characterize each stage in the differentiation process, (2) that gene expression can be efficiently depleted throughout the differentiation time course using shRNAs expressed from lentiviruses and (3) that the nuclear hormone receptor HNF4A is essential for specification of human hepatic progenitor cells by establishing the expression of the network of transcription factors that controls the onset of hepatocyte cell fate.


Screening for colorectal cancer. Issues for primary care physicians.
Fecal occult blood testing, proctosigmoidoscopy, and digital rectal examination have been recommended as screening tests for colorectal cancer in asymptomatic people. This article evaluates the advisability of recommendations by applying accepted principles of screening to the case of colorectal cancer.


The dopamine D1 receptor is expressed and induces CREB phosphorylation and MUC5AC expression in human airway epithelium.
Dopamine receptors comprise two subgroups, Gs protein-coupled "D1-like" receptors (D1, D5) and Gicoupled "D2-like" receptors (D2, D3, D4). In airways, both dopamine D1 and D2 receptors are expressed on airway smooth muscle and regulate airway smooth muscle force. However, functional expression of the dopamine D1 receptor has never been identified on airway epithelium. Activation of Gs-coupled receptors stimulate adenylyl cyclase leading to cyclic AMP (cAMP) production, which is known to induce mucus overproduction through the cAMP response element binding protein (CREB) in airway epithelial cells. We questioned whether the dopamine D1 receptor is expressed on airway epithelium, and whether it promotes CREB phosphorylation and MUC5AC expression. We evaluated the protein expression of the dopamine D1 receptor on native human airway epithelium and three sources of cultured human airway epithelial cells including primary cultured airway epithelial cells, the bronchial epithelial cell line (16HBE14o-), and the pulmonary mucoepidermoid carcinoma cell line (NCI-H292) using immunohistochemistry and immunoblotting. To characterize the stimulation of cAMP through the dopamine D1 receptor, 16HBE14o- cells and NCI-H292 cells were treated with dopamine or the dopamine D1 receptor agonists (SKF38393 or A68930) before cAMP measurements. The phosphorylation of CREB by A68930 in both 16HBE14o- and NCI-H292 cells was measured by immunoblot. The effect of dopamine or A68930 on the expression of MUC5AC mRNA and protein in NCI-H292 cells was evaluated by real-time PCR and immunofluorescence staining, respectively. The dopamine D1 receptor protein was detected in native human airway epithelium and three sources of cultured human airway epithelial cells. Dopamine or the dopamine D1-like receptor agonists stimulated cAMP production in 16HBE14o- cells and NCI-H292 cells, which was reversed by the selective dopamine D1-like receptor antagonists (SCH23390 or SCH39166). A68930 significantly increased phosphorylation of CREB in both 16HBE14o- and NCI-H292 cells, which was attenuated by the inhibitors of PKA (H89) and MEK (U0126). Expression of MUC5AC mRNA and protein were also increased by either dopamine or A68930 in NCI-H292 cells. These results suggest that the activation of the dopamine D1 receptor on human airway epithelium could induce mucus overproduction, which could worsen airway obstructive symptoms.


Screening for mammalian neural genes via fluorescence-activated cell sorter purification of neural precursors from Sox1-gfp knock-in mice.
The transcription factor Sox1 is the earliest and most specific known marker for mammalian neural progenitors. During fetal development, Sox1 is expressed by proliferating progenitor cells throughout the central nervous system and in no tissue but the lens. We generated a reporter mouse line in which egfp is inserted into the Sox1 locus. Sox1GFP animals faithfully recapitulate the expression of the endogenous gene. We have used the GFP reporter to purify neuroepithelial cells by fluorescence-activated cell sorting from embryonic day 10.5 embryos. RNAs prepared from Sox1GFP+ and Sox1GFP- embryo cells were then used to perform a pilot screen of subtracted cDNAs prepared from differentiating embryonic stem cells and arrayed on a glass chip. Fifteen unique differentially expressed genes were identified, all previously associated with fetal or adult neural tissue. Whole mount in situ hybridization against two genes of previously unknown embryonic expression, Lrrn1 and Musashi2, confirmed the selectivity of this screen for early neuroectodermal markers.


Corneal epithelial changes in a patient treated with belantamab mafodotin.
The aim of the study is to report a case of corneal epithelial changes in a patient with refractory multiple myeloma (MM) who was treated with belantamab mafodotin (BM). A 55-year-old man diagnosed with refractory MM was referred to our hospital for treatment with BM, an antibody-drug conjugate targeting B-cell maturation antigen. After 33 days of treatment, the patient experienced a bilateral reduction in visual acuity (VA), along with dry eye symptoms such as itchy eyes and a sensation of having a foreign body. Slit-lamp examination revealed the presence of diffuse microcystic epithelial changes throughout the cornea. BM treatment was discontinued by the oncologist. Sixty days after stopping belantamab, VA gradually improved and the microcystic epithelial alterations progressively diminished. Ninety days after discontinuation of therapy, only a few microcystic epithelial alterations remained, and the patient had 20/20 VA in both eyes. While BM is an effective therapy for refractory MM, corneal epithelial changes are among the most common side effects of this treatment. Close collaboration between ophthalmologists and oncologists is crucial for assessing ocular adverse effects and tailoring treatment accordingly.


Racial/Ethnic Differences in Provider-Engaged Religious Belief Discussions with Older Adults at the End of Life.
The purpose of this study was to determine racial/ethnic differences in provider-engaged religious belief discussion with older adults in the final month of life. Data were derived from the combined 2012 to 2020 National Health and Aging Trends Study. Chi-square tests were used for bivariate analysis, and a binary logistic regression model was used to test the association between race/ethnicity and provider-engaged religious belief discussions at the end of life. After controlling for other explanatory factors during the analysis, results showed that providers were less likely to have religious belief discussions with Black and Hispanic older adults compared to Whites. The results of this study point to a significant gap in knowledge among healthcare providers whose diversity training may not be inclusive of religious/spiritual cultural humility. Implications for research, policy, and practice are provided.


Spatial filtering velocimetry of objective speckles for measuring out-of-plane motion.
This paper analyzes the dynamics of objective laser speckles as the distance between the object and the observation plane continuously changes. With the purpose of applying optical spatial filtering velocimetry to the speckle dynamics, in order to measure out-of-plane motion in real time, a rotational symmetric spatial filter is designed. The spatial filter converts the speckle dynamics into a photocurrent with a quasi-sinusoidal response to the out-of-plane motion. The spatial filter is here emulated with a CCD camera, and is tested on speckles arising from a real application. The analysis discusses the selectivity of the spatial filter, the nonlinear response between speckle motion and observation distance, and the influence of the distance-dependent speckle size. Experiments with the emulated filters illustrate performance and potential applications of the technology.


Identification of a second GTP-bound magnesium ion in archaeal initiation factor 2.
Eukaryotic and archaeal translation initiation processes involve a heterotrimeric GTPase e/aIF2 crucial for accuracy of start codon selection. In eukaryotes, the GTPase activity of eIF2 is assisted by a GTPase-activating protein (GAP), eIF5. In archaea, orthologs of eIF5 are not found and aIF2 GTPase activity is thought to be non-assisted. However, no in vitro GTPase activity of the archaeal factor has been reported to date. Here, we show that aIF2 significantly hydrolyses GTP in vitro. Within aIF2γ, H97, corresponding to the catalytic histidine found in other translational GTPases, and D19, from the GKT loop, both participate in this activity. Several high-resolution crystal structures were determined to get insight into GTP hydrolysis by aIF2γ. In particular, a crystal structure of the H97A mutant was obtained in the presence of non-hydrolyzed GTP. This structure reveals the presence of a second magnesium ion bound to GTP and D19. Quantum chemical/molecular mechanical simulations support the idea that the second magnesium ion may assist GTP hydrolysis by helping to neutralize the developing negative charge in the transition state. These results are discussed in light of the absence of an identified GAP in archaea to assist GTP hydrolysis on aIF2.


Genotyping of a case of tyrosinaemia type I with normal level of succinylacetone in amniotic fluid.
Tyrosinaemia type I is caused by a deficiency of fumarylacetoacetate hydrolase and mainly affects the liver. This disease is characterized by the presence of a high level of succinylacetone. This metabolite has been used for prenatal diagnosis from amniotic fluid samples. One case with a normal level of succinylacetone in amniotic fluid has recently been described (Grenier et al., 1996). Here, we report that this patient is a compound heterozygote for two known mutations: E364X and IVS6-1g-->t. The low level of succinylacetone cannot be explained by these mutations.


Provider, veteran, and family perspectives on family education in Veterans Affairs community-based outpatient facilities.
The Veterans Affairs (VA) healthcare system is dedicated to providing high-quality mental health services to all veterans, including the nearly 40% of enrolled veterans living in rural areas. Family education programs regarding mental illness and posttraumatic stress disorder, mandated for delivery in all VA medical centers and some community-based outpatient clinics (CBOCs), have been developed and provided primarily in large, urban medical centers. This qualitative investigation involved interviews with CBOC providers and veterans and families who live in rural areas and/or seek care in CBOCs to ascertain their perceptions of the benefits, feasibility, structural and cultural barriers, and logistical preferences regarding family education. The perspectives and concerns that emerged in these interviews were combined with expert knowledge to identify the resources and considerations a VAMC would want to address when translating and implementing similar programming into CBOCs. Although institutional, logistic, and attitudinal challenges were described, all three stakeholder groups endorsed the need for family education, did not see the barriers as insurmountable, and provided creative solutions. Administrators and CBOC clinicians may benefit by anticipating and problem solving around the key issues raised when developing family programming.


[Lymphoepithelial cyst of the pancreas. A case report].
Lymphoepithelial cyst of the pancreas is a benign and rare pathology. Its histogenesis is still unknown. The diagnosis is difficult to establish before surgery. We report a new case of a 20-year-old woman admitted for abdominal pain and vomiting. Radiologic investigations described a multilocular cystic tumor of the tail of the pancreas. The patient underwent a left pancreatectomy with splenectomy. Histologic investigations revealed pancreatic cysts lined by squamous epithelium surrounded by dense lymphoid tissue. The diagnosis of lymphoepithelial cyst of the pancreas was done.


Disappearance of glucagonoma rash after surgical resection, but not during dietary normalization of serum amino acids.
Hypoaminoacidemia and skin rash are features of the glucagonoma syndrome. A glucagonoma patient with earlier insulin treated, noninsulin-dependent diabetes, and functional liver metastases, 3 yr postresection of pancreatic tumor, was treated with a high protein diet for 2 wk, then switched to a high carbohydrate diet for 3 wk followed by 3 wk on high protein diet, which continued for 3 months with additional carbohydrate. While on the high protein diet urine nitrogen indicated frank retention and total plasma amino acid levels normalized each time. Plasma amino acid decreased again after 1 wk on the high carbohydrate diet. Skin rash varied irrespective of amino acid levels, but cleared 4 days after resection of metastases. However, total amino acid did not reach normal levels on a conventional diet 3 wk postsurgery, but were normal 6 wk later. A high protein diet can normalize plasma amino acids and allow nitrogen retention in glucagonoma, apparently overriding the gluconeogenic drive of the high circulating glucagon levels. The skin rash may not be only attributed to hypoaminoacidemia.


Defining extraordinary care.
Criteria-based protocols are an ethically appropriate strategy to addressing issues such as allocation of scarce resources; however, protocols can conflict with the ethical ideal of respect for individual autonomy. This article illustrates the conflict that can occur between individual patient situations and standardized criteria-based protocols. Furthermore, ordinary care and treatment outlined in criteria-based protocols can become extraordinary in some cases. The case study presented illustrates why it is important to distinguish between ordinary and extraordinary care in some situations. The distinction between the two is described herein.


Benzodiazepines in Miami-Dade County, Florida driving under the influence (DUI) cases (1995-1998) with emphasis on Rohypnol: GC-MS confirmation, patterns of use, psychomotor impairment, and results of Florida legislation.
Benzodiazepines are central nervous system depressant drugs often detected in biological samples from driving under the influence (DUI) offenders. They are associated with marked psychomotor impairment and represent up to 20% of all Miami-Dade County, Florida DUI urine samples analyzed in our laboratory annually. Flunitrazepam emerged in the mid-1990s as an illegal drug in the U.S. that was predominantly abused recreationally and associated with sexual assaults. Immunoassays for benzodiazepines do not discriminate between different benzodiazepines, and certain metabolites, such as 7-aminoflunitrazepam, react poorly with immunoassay reagents. A simple and sensitive method for the detection and quantitation of major benzodiazepines and metabolites by gas chromatography with mass selective detection is presented. This method was used to confirm benzodiazepines in general and flunitrazepam in particular. Data collected over a three-and-a-half-year period are summarized. Whereas flunitrazepam was present in up to 10% of DUI cases in 1995 and 1996 and had fast become the most frequently encountered benzodiazepine in Miami-Dade County DUI-related urine samples, a dramatic drop in case numbers followed the legal reclassification of the drug as a Schedule I substance in Florida in February 1997. Flunitrazepam was often used alone or in combination with cannabis and cocaine. A recent rise in clonazepam cases coincides with the decrease in flunitrazepam confirmation and may indicate a new trend in the abuse of benzodiazepines in South Florida.


Altered Spatial Organization of Dynamic Functional Network Associates With Deficient Sensory and Perceptual Network in Schizophrenia.
Schizophrenia is currently thought as a disorder with dysfunctional communication within and between sensory and cognitive processes. It has been hypothesized that these deficits mediate heterogeneous and comprehensive schizophrenia symptomatology. In this study, we investigated as to how the abnormal dynamic functional architecture of sensory and cognitive networks may contribute to these symptoms in schizophrenia. We calculated a sliding-window-based dynamic functional connectivity strength (FCS) and amplitude of low-frequency fluctuation (ALFF) maps. Then, using group-independent component analysis, we characterized spatial organization of dynamic functional network (sDFN) across various time windows. The spatial architectures of FCS/ALFF-sDFN were similar with traditional resting-state functional networks and cannot be accounted by length of the sliding window. Moreover, schizophrenic subjects demonstrated reduced dynamic functional connectivity (dFC) within sensory and perceptual sDFNs, as well as decreased connectivity between these sDFNs and high-order frontal sDFNs. The severity of patients' positive and total symptoms was related to these abnormal dFCs. Our findings revealed that the sDFN during rest might form the intrinsic functional architecture and functional changes associated with psychotic symptom deficit. Our results support the hypothesis that the dynamic functional network may influence the aberrant sensory and cognitive function in schizophrenia, further highlighting that targeting perceptual deficits could extend our understanding of the pathophysiology of schizophrenia.


Cooling, heating, generating power, and recovering waste heat with thermoelectric systems.
Thermoelectric materials are solid-state energy converters whose combination of thermal, electrical, and semiconducting properties allows them to be used to convert waste heat into electricity or electrical power directly into cooling and heating. These materials can be competitive with fluid-based systems, such as two-phase air-conditioning compressors or heat pumps, or used in smaller-scale applications such as in automobile seats, night-vision systems, and electrical-enclosure cooling. More widespread use of thermoelectrics requires not only improving the intrinsic energy-conversion efficiency of the materials but also implementing recent advancements in system architecture. These principles are illustrated with several proven and potential applications of thermoelectrics.


Incidence of syphilis in Greenland 2010-2014: the beginning of a new epidemic?
To estimate the incidence of syphilis from 2010 to 2014 and to assess whether contact tracing has been performed. Observational cross-sectional study. Data on reported cases were collected from the national register of the chief medical officer in Greenland. Unreported cases were found by searching the electronic medical record system for patients who had received an electronic prescription of benzathine penicillin, doxycycline or tetracycline. Medical records were reviewed to verify the diagnosis of syphilis and to evaluate if contact tracing had been performed. Ninety-four cases of syphilis (51 males and 43 females) with a median age of 27 years (20-40) were included. The incidence of syphilis in Greenland has increased from zero cases in 2010 to 95.7 per 100,000 inhabitants in 2014 affecting mainly young people. Contact tracing was performed in 80.9% (76/94) of the cases. Syphilis has re-occurred in Greenland and a new epidemic may be underway. Sustained awareness of treatment, contact tracing, monitoring and preventive initiatives are desirable.


Herpes simplex virus type 1 serum neutralizing antibody titers increase during latency in rabbits latently infected with latency-associated transcript (LAT)-positive but not LAT-negative viruses.
The herpes simplex virus type 1 (HSV-1) latency-associated transcript (LAT) gene is essential for efficient spontaneous reactivation in the rabbit ocular model of HSV-1 latency and reactivation. LAT is also the only viral gene abundantly expressed during latency. Rabbits were ocularly infected with the wild-type HSV-1 strain McKrae or the McKrae-derived LAT null mutant dLAT2903. Serum neutralizing antibody titers were determined at various times during acute and latent infection. The neutralizing antibody titers induced by both viruses increased and were similar throughout the first 45 days after infection (P > 0.05). However, by day 59 postinfection (approximately 31 to 45 days after latency had been established), the neutralizing antibody titers induced by wild-type virus and dLAT2903 diverged significantly (P = 0.0005). The dLAT2903-induced neutralizing antibody titers decreased, while the wild-type virus-induced neutralizing antibody titers continued to increase. A rescuant of dLAT2903, in which spontaneous reactivation was fully restored, induced wild-type neutralizing antibody levels on day 59 postinfection. A second LAT mutant with impaired spontaneous reactivation had neutralizing antibody levels comparable to those of dLAT2903. In contrast to the results obtained in rabbits, in mice, neutralizing antibody titers did not increase over time during latency with any of the viruses. Since LAT is expressed in both rabbits and mice during latency, the difference in neutralizing antibody titers between these animals is unlikely to be due to expression of a LAT protein during latency. In contrast, LAT-positive (LAT(+)), but not LAT-negative (LAT(-)), viruses undergo efficient spontaneous reactivation in rabbits, while neither LAT(+) nor LAT(-) viruses undergo efficient spontaneous reactivation in mice. Thus, the increase in neutralizing antibody titers in rabbits latently infected with LAT(+) viruses may have been due to continued restimulation of the immune system by spontaneously reactivating virus.


Targeted delivery of genistein for pancreatic cancer treatment using hyaluronic-coated cubosomes bioactivated with frankincense oil.
Pancreatic cancer is an aggressive malignancy that remains a major cause of cancer-related deaths. Research for innovative anticancer therapeutic options is thus imperative. In this regard, phytotherapeutics offer great promise as efficient treatment modalities, especially leveraging nanodrug delivery. Herein, we innovatively coloaded the flavonoid genistein (Gen) and frankincense essential oil (FO) within cubosomes, which were then coated with the bioactive ligand hyaluronic acid (HA/Gen-FO-Cub) for active-targeting of pancreatic cancer. The novel HA/Gen-FO-Cub displayed optimum nanosize (198.2 ± 4.5 nm), PDI (0.27 ± 0.01), zeta-potential (-34.7 ± 1.2 mV), Gen entrapment (99.3 ± 0.01 %), and controlled Gen release (43.7 ± 1.2 % after 120 h). HA/Gen-FO-Cub exerted selective anticancer activity on pancreatic cancer cells (PANC-1; 8-fold drop in IC50), cellular uptake and anti-migratory effect compared to Gen solution. HA/Gen-FO-Cub revealed prominent cytocompatibility (100 ± 5.9 % viability of human dermal fibroblast). Moreover, HA/Gen-FO-Cub boosted the in vivo anticancer activity of Gen in an orthotopic cancer model, affording tumor growth suppression (2.5-fold drop) and downregulation of NFκB and VEGF (2.9- and 1.8-fold decrease, respectively), compared to Gen suspension. Antimetastatic efficacy and Bcl-2-downexpression was histologically confirmed. Our findings demonstrate the promising anticancer aptitude of HA/Gen-FO-Cub as an effective phytotherapeutic nanodelivery system for pancreatic cancer therapy.


Cytotoxicity of the Vibrio vulnificus MARTX toxin effector DUF5 is linked to the C2A subdomain.
The multifunctional-autoprocessing repeats-in-toxin (MARTX) toxins are bacterial protein toxins that serve as delivery platforms for cytotoxic effector domains. The domain of unknown function in position 5 (DUF5) effector domain is present in at least six different species' MARTX toxins and as a hypothetical protein in Photorhabdus spp. Its presence increases the potency of the Vibrio vulnificus MARTX toxin in mouse virulence studies, indicating DUF5 directly contributes to pathogenesis. In this work, DUF5 is shown to be cytotoxic when transiently expressed in HeLa cells. DUF5 localized to the plasma membrane dependent upon its C1 domain and the cells become rounded dependent upon its C2 domain. Both full-length DUF5 and the C2 domain caused growth inhibition when expressed in Saccharomyces cerevisiae. A structural model of DUF5 was generated based on the structure of Pasteurella multocida toxin facilitating localization of the cytotoxic activity to a 186 amino acid subdomain termed C2A. Within this subdomain, an alanine scanning mutagenesis revealed aspartate-3721 and arginine-3841 as residues critical for cytotoxicity. These residues were also essential for HeLa cell intoxication when purified DUF5 fused to anthrax toxin lethal factor was delivered cytosolically. Thermal shift experiments indicated that these conserved residues are important to maintain protein structure, rather than for catalysis. The Aeromonas hydrophila MARTX toxin DUF5(Ah) domain was also cytotoxic, while the weakly conserved C1-C2 domains from P. multocida toxin were not. Overall, this study is the first demonstration that DUF5 as found in MARTX toxins has cytotoxic activity that depends on conserved residues in the C2A subdomain.


Labeling stem cells with ferumoxytol, an FDA-approved iron oxide nanoparticle.
Stem cell based therapies offer significant potential for the field of regenerative medicine. However, much remains to be understood regarding the in vivo kinetics of transplanted cells. A non-invasive method to repetitively monitor transplanted stem cells in vivo would allow investigators to directly monitor stem cell transplants and identify successful or unsuccessful engraftment outcomes. A wide range of stem cells continues to be investigated for countless applications. This protocol focuses on 3 different stem cell populations: human embryonic kidney 293 (HEK293) cells, human mesenchymal stem cells (hMSC) and induced pluripotent stem (iPS) cells. HEK 293 cells are derived from human embryonic kidney cells grown in culture with sheared adenovirus 5 DNA. These cells are widely used in research because they are easily cultured, grow quickly and are easily transfected. hMSCs are found in adult marrow. These cells can be replicated as undifferentiated cells while maintaining multipotency or the potential to differentiate into a limited number of cell fates. hMSCs can differentiate to lineages of mesenchymal tissues, including osteoblasts, adipocytes, chondrocytes, tendon, muscle, and marrow stroma. iPS cells are genetically reprogrammed adult cells that have been modified to express genes and factors similar to defining properties of embryonic stem cells. These cells are pluripotent meaning they have the capacity to differentiate into all cell lineages. Both hMSCs and iPS cells have demonstrated tissue regenerative capacity in-vivo. Magnetic resonance (MR) imaging together with the use of superparamagnetic iron oxide (SPIO) nanoparticle cell labels have proven effective for in vivo tracking of stem cells due to the near microscopic anatomical resolution, a longer blood half-life that permits longitudinal imaging and the high sensitivity for cell detection provided by MR imaging of SPIO nanoparticles. In addition, MR imaging with the use of SPIOs is clinically translatable. SPIOs are composed of an iron oxide core with a dextran, carboxydextran or starch surface coat that serves to contain the bioreactive iron core from plasma components. These agents create local magnetic field inhomogeneities that lead to a decreased signal on T2-weighted MR images. Unfortunately, SPIOs are no longer being manufactured. Second generation, ultrasmall SPIOs (USPIO), however, offer a viable alternative. Ferumoxytol (FerahemeTM) is one USPIO composed of a non-stoichiometric magnetite core surrounded by a polyglucose sorbitol carboxymethylether coat. The colloidal, particle size of ferumoxytol is 17-30 nm as determined by light scattering. The molecular weight is 750 kDa, and the relaxivity constant at 2T MRI field is 58.609 mM(-1) sec(-1) strength. Ferumoxytol was recently FDA-approved as an iron supplement for treatment of iron deficiency in patients with renal failure. Our group has applied this agent in an "off label" use for cell labeling applications. Our technique demonstrates efficient labeling of stem cells with ferumoxytol that leads to significant MR signal effects of labeled cells on MR images. This technique may be applied for non-invasive monitoring of stem cell therapies in pre-clinical and clinical settings.


Effect of sodium naproxen on inflammatory response induced by anterior chamber paracentesis in the rabbit.
This study evaluated the effect of sodium naproxen (a reversible competitive inhibitor of cyclo-oxygenase) and phenylephrine (a mydriatic alpha-adrenergic agent) eye drops in maintaining atropine mydriasis in the rabbit after paracentesis. Moreover, to assess the influence of these treatments on vascular and cellular inflammatory responses in the rabbit eye, several biochemical parameters were considered. Anterior chamber paracentesis significantly reduced atropine-induced mydriasis and a parallel elevation of proteins, polymorphonuclear leucocytes (PMNs), prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) levels in the secondary aqueous humour (obtained 120 min later) was observed. A significant increase in PMNs in the aqueous humour and a parallel increase in myeloperoxidase activity, a measure of PMN infiltration, in the iris-ciliary body were detected. Atropine-induced mydriasis was maintained in rabbits treated with either sodium naproxen or phenylephrine eye drops. However, only in the former group were the inflammatory parameters significantly reduced, with the exception of aqueous LTB4 levels. The inhibition of the protein influx in the aqueous humour and of the miosis produced by sodium naproxen can be related to the high drug levels in the aqueous humour that were effective in inhibiting the cyclo-oxygenase pathway of arachidonic acid metabolism, whereas the effects on PMN infiltration appear to be independent of significant release of the potent chemotactic agent LTB4, synthesized via the 5-lipoxygenase pathway.


Association of foetal size and sex with porcine foeto-maternal interface integrin expression.
Integrins regulate adhesion at the foeto-maternal interface by interacting with secreted phosphoprotein 1 (SPP1) and fibronectin (FN). It is hypothesised that impaired foetal growth of 'runt' piglets is linked to altered integrin signalling at the foeto-maternal interface. Placental and endometrial samples associated with the lightest and closest to mean litter weight (CTMLW) (gestational day (GD18, 30, 45, 60 and 90), of both sex (GD30, 45, 60 and 90) (n = 5-8 litters/GD), Large White × Landrace conceptuses or foetuses were obtained. The mRNA expression of the integrin subunits (ITG) ITGA2, ITGAV, ITGB1, ITGB3, ITGB5, ITGB6, ITGB8, SPP1 and FN was quantified by qPCR. Temporal changes in mRNA expression were observed, with different profiles in the two tissues. Endometrial ITGB1 (P ≤ 0.05, GD45) and SPP1 (P ≤ 0.05, all GD combined and GD60) expression was decreased in samples supplying the lightest compared to the CTMLW foetuses. Placentas supplying female foetuses had decreased expression of ITGB6 (GD45, P ≤ 0.05) and FN (GD90, P ≤ 0.05) compared to those supplying male foetuses. Endometrial samples supplying females had increased ITGB3 (P ≤ 0.05, GD60) and FN (P ≤ 0.05, GD30) expression and decreased SPP1 (P ≤ 0.05, GD60) expression compared to male foetuses. Correlations between mean within-gilt mRNA expression and percentage prenatal survival, number of live foetuses or conceptuses and percentage male foetuses were observed. This study has highlighted novel and dynamic associations between foetal size, sex and integrin subunit mRNA expression at the porcine foeto-maternal interface. Further studies should be performed to improve the understanding of the mechanisms behind these novel findings.


Impact of withdrawal of the analgesic Co-proxamol on nonfatal self-poisoning in the UK.
In early 2005 the UK Committee on Safety of Medicines (CSM) announced gradual withdrawal of the analgesic co-proxamol because of its adverse benefit/safety ratio, especially its use for intentional and accidental fatal poisoning. Prescriptions of co-proxamol were reduced in the 3-year withdrawal phase (2005 to 2007) following the CSM announcement. To assess the impact of the CSM announcement in January 2005 to withdraw co-proxamol on nonfatal self-poisoning with co-proxamol and other analgesics. Interrupted time series analysis of general hospital presentations for nonfatal self-poisoning (five hospitals in three centers in England), comparing the 3-year withdrawal period 2005-2007 with 2000-2004. A marked reduction in the number of episodes of nonfatal self-poisoning episodes involving co-proxamol was found following the CSM announcement (an estimated 62% over the period 2005 to 2007 compared to 2000 to 2004). There was no evidence of an increase in nonfatal self-poisoning episodes involving other analgesics (co-codamol, codeine, co-dydramol, dihydrocodeine, and tramadol) in relation to the CSM announcement over the same period, nor a change in the number of all episodes of self-poisoning. Data were from three centers only. The impact of the policy appears to have reduced nonfatal self-poisoning with co-proxamol without significant substitution with other analgesics. This finding is in keeping with that for suicide.


Safety and Dose Escalation Study of Intravenous Zinc Supplementation in Pediatric Critical Illness.
Critically ill children have low plasma zinc (pZn), correlating with organ failure. Since Zn influences inflammation, immune function, and glucose control, Zn supplementation is a plausible therapeutic modality. We sought to determine a safe dose of intravenous (IV) Zn to restore pZn in critically ill children. Stepwise dose escalation study of IV Zn supplementation at a tertiary children's hospital. All children (<10 years) admitted to the pediatric intensive care unit with a Pediatric Risk of Mortality III score >5, or ≥1 new organ failure were eligible. After consent, patients were sequentially enrolled into 4 dosing groups: (1) no zinc, (2) Zn250: 250 mcg/kg/d ZnSO4, (3) Zn500: 500 mcg/kg/d ZnSO4, or (4) Zn750: 750 mcg/kg/d ZnSO4 ZnSO4 was administered 3 times daily for 7 days. pZn was measured at baseline, end of first ZnSO4 infusion, 1 hour postinfusion, and 7 hours postinfusion on day 1, then daily through days 2-7. Interleukin-6 (IL-6), C-reactive protein (CRP), and lymphocyte subsets were measured on days 1 and 3. Glucose was measured 3 times daily for 7 days. Twenty-four patients were enrolled. Baseline demographics were similar among groups. Baseline pZn was low in all patients (mean [SD], 41.8 [16.0] mcg/dL). pZn increased over the study period in supplemented groups; however, mean pZn in the Zn750 group exceeded the 50th percentile. pZn was not associated with IL-6, CRP, or lymphocyte subsets among groups. Degree of hyperglycemia did not differ among groups. No patient had a study-related adverse event. IV zinc supplementation at 500 mcg/kg/d restores pZn to near the 50th percentile and is well tolerated.


Effects of diclofenac-sodium and salicylate, carticaine, lidocaine and procaine in the cadmium- and heat-induced hemolysis.
In the present study we examined the utility, sensitivity and specificity of the cadmium- and heat-induced hemolysis, investigating the effects of diclofenac, salicylate and lidocaine, carticaine and procaine on the rabbit erythrocyte membrane. In the cadmium-induced hemolysis, which has not been reported, as far as we know, as a test model for local anesthetics, the studied local anesthetics have, to a large extent, protective effects on the erythrocyte membranes, probably based on an osmotic action at drug concentrations of 10(-4) to 10(-2) M. On the other hand, these local anesthetics amplify the heat-induced hemolysis to a variable extent. The studied antirheumatic drugs showed in the cadmium-induced hemolysis that they cannot have the same binding sites as the local anesthetics investigated. In the cadmium-induced hemolysis, the reaction products of the antirheumatic drugs with the membrane protein presumably cannot cause an increase in surface area/volume ratio of the erythrocytes, which is generally regarded as a cause for stabilization. This interpretation could be an explanation for the increase of the cadmium-induced hemolysis by the antirheumatic drugs studied. On the other hand, the antirheumatic drugs inhibited to a variable extent the heat-induced hemolysis.


[Lymphadenectomy for thyroid and lymph node carcinomas].
In general, primary surgery of thyroid carcinoma should consist of total thyroidectomy and lymph node dissection of the cervicocentral compartment. Exceptions are cases of papillary microcarcinoma and prophylactic surgery due to multiple type 2A endocrine neoplasia. Lymph node dissection beyond the cervicocentral compartment also should be compartment-oriented. It is generally indicated if lymph node metastases have been proven. Concerning clinically proven medullary thyroid carcinoma, bilateral cervicolateral lymph node dissection is generally indicated, since lymph node metastases may be missed preoperatively but are often found histologically. In patients with parathyroid carcinoma, en bloc ipsilateral cervicocentral lymph node dissection should be performed in addition to parathyroidectomy and hemithyroidectomy. Lymph node dissection should always be performed systematically, since lymph node metastases may be missed both clinically and by imaging techniques.


Treatment of Intestinal Inflammation With Epicutaneous Immunotherapy Requires TGF-β and IL-10 but Not Foxp3+ Tregs.
Background: Inflammatory bowel disease (IBD) involves an increase in T effector cells in the intestines that disrupts the normal balance with T regulatory cells (Tregs). A therapy that restores this balance has the potential to treat IBD. We have shown that epicutaneous exposure to OVA induces Tregs that are able to induce tolerance. The Tregs also migrate to the intestines where they alleviate colitis in mice, demonstrating the potential for skin induced Tregs to treat intestinal inflammation. We investigated the role of Foxp3, IL-10, and TGF-β in the suppression of colitis by epicutaneous immunotherapy (ET). Methods: RAG1-/- mice were transferred with CD4+CD45RBhi T cells from wild type mice to induce colitis. To determine whether Foxp3+ Tregs, IL-10-, or TGF-β-producing Tregs were necessary, Foxp3-DTR, IL-10-/-, or CD4-dnTGFBRII mice were immunized with OVA and OVA TCR enriched T cells were added. As control groups, some mice were given OVA TCR enriched T cells from wild type mice or no OVA TCR enriched T cells. Half of the mice in each group were then exposed on the skin to Viaskin patches containing OVA weekly for 3 weeks. Mice given OVA TCR enriched T cells from Foxp3-DTR mice were given diphtheria toxin (DT) or not in addition to ET. Mice were assessed for weight loss, colon length, colonic cytokine production, and histological inflammation. Results: ET, after injection with OVA TCR enriched T cells derived from wild type mice, prevented weight loss, decreased colonic inflammatory cytokine production and histological colitis. ET in the absence of the OVA TCR enriched T cells did not alleviate colitis. ET, after injection with OVA TCR enriched T cells derived from Foxp3-DTR mice, prevented weight loss, decreased colonic inflammatory cytokine production, and histological colitis. Ablation with DT did not impair the ability of ET to alleviate colitis. ET failed to alleviate colitis when OVA TCR enriched T cells were derived from IL-10-/- or CD4-dnTGFBRII mice. Conclusions: ET through induction of Tregs, which produce IL-10 and TGF-β, could be a promising treatment for IBD.


Topographical distribution of barrier function in cervico-cephalic arteries of dog. Major cerebral arteries possess definite barrier function?
Topographic distribution of barrier function in normal canine cervico-cephalic arteries was studied using horseradish peroxidase (HRP) and Evans blue as tracers. The carotid sinus of the internal carotid artery (ICA) was conspicuously permeable to HRP when compared to other areas of major cervical arteries. The cavernous portion of the ICA also showed prominent permeation of HRP, especially through the outer surface, which is covered with venous endothelial cells. On the luminal side of the cavernous portion of the ICA, barrier deficiency was noted at angulated segments such as the carotid siphon. Intracranial segments of both ICA and vertebral arteries demonstrated incomplete barrier function of the first 1 to 4 mm from the origin of the intradural segments. These areas were considered to be transitional sites in barrier function between extra- and intracranial arteries. Focal, but definite, barrier disruption was also noted at the distal ends of the ICA and other arterial branching sites of major intracranial arteries. While opening of the interendothelial junctions was considered to be one of the mechanisms causing increased permeability in the cavernous ICA, the mechanisms for the permeation of HRP into the major cerebral arteries could not be confirmed ultrastructurally.


Synovial calprotectin: a potential biomarker to exclude a prosthetic joint infection.
Recently, several synovial biomarkers have been introduced into the algorithm for the diagnosis of a prosthetic joint infection (PJI). Alpha defensin is a promising biomarker, with a high sensitivity and specificity, but it is expensive. Calprotectin is a protein that is present in the cytoplasm of neutrophils, is released upon neutrophil activation and exhibits anti-microbial activity. Our aim, in this study, was to determine the diagnostic potential of synovial calprotectin in the diagnosis of a PJI. In this pilot study, we prospectively collected synovial fluid from the hip, knee, shoulder and elbow of 19 patients with a proven PJI and from a control group of 42 patients who underwent revision surgery without a PJI. PJI was diagnosed according to the current diagnostic criteria of the Musculoskeletal Infection Society. Synovial fluid was centrifuged and the supernatant was used to measure the level of calprotectin after applying a lateral flow immunoassay. The median synovial calprotectin level was 991 mg/L (interquartile range (IQR) 154 to 1787) in those with a PJI and 11 mg/L (IQR 3 to 29) in the control group (p < 0.0001). Using a cut-off value of 50 mg/L, this level showed an excellent diagnostic accuracy, with an area under the curve of 0.94. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) was 89%, 90%, 81% and 95% respectively. The NPV was 97% in the nine patients with a chronic PJI. Synovial calprotectin may be a valuable biomarker in the diagnosis of a PJI, especially in the exclusion of an infection. With a lateral flow immunoassay, a relatively rapid quantitative diagnosis can be made. The measurement is cheap and is easy to use. Cite this article: Bone Joint J 2017;99-B:660-5.


[Use of the artificial endocrine pancreas in the clinic].
Via the artificial endocrine pancreas--apparatus "BIOSTATOR" 100 patients were studied and treated, namely: 90 with diabetes mellitus, 7 with hyperinsulinism and 3 with obesity. The 24-h insulin needs of the diabetics with insulin-dependent and non-insulin dependent with secondary exhaustion type of diabetes, were determined. The following subcutaneous insulin dosage was determined, depending on the insulin amount spent during the 24-h biostator control. It was established that the necessary reduction of the intravenously administered insulin is 30% on the average with the passing over to subcutaneous regime. The indices of lipid metabolism were also studied as well as numerous hormones with which the carbohydrate compensation leads to a change. The problem of the "morning" phenomenon is discussed.


Time course and time-distance relationships for surround suppression in macaque V1 neurons.
Iso-orientation surround suppression is a powerful form of visual contextual modulation in which a stimulus of the preferred orientation of a neuron placed outside the classical receptive field (CRF) of the neuron suppresses the response to stimuli within the CRF. This suppression is most often attributed to orientation-tuned signals that propagate laterally across the cortex, activating local inhibition. By studying the temporal properties of surround suppression, we have uncovered characteristics that challenge standard notions of surround suppression. We found that the latency of suppression depended on its strength. Across cells, strong suppression arrived on average 30 msec earlier than weak suppression, and suppression sometimes arrived faster than the excitatory CRF response. We compared the relative latency of CRF response onset and offset with the relative latency of suppression onset and offset. Response onset was delayed relative to response offset in the CRF but not in the surround. This is not the expected result if neurons targeted by suppression are like those that generate it. We examined the time course of suppression as a function of distance of the surround stimulus from the CRF and found that suppression was predominantly sustained for nearby stimuli and predominantly transient for distant stimuli. By comparing the latency of suppression for nearby and distant stimuli, we found that orientation-tuned suppression could effectively propagate across 6 - 8 mm of cortex at approximately 1 m/sec. This is considerably faster than expected for horizontal cortical connections previously implicated in surround suppression. We offer refinements to circuits for surround suppression that account for these results and describe how feedback from cells with large CRFs can account for the rapid propagation of suppression within V1.