Similar serum levels of IL-6 and its soluble receptors in patients with HCV-related arthritis and rheumatoid arthritis: a pilot study.

The high serum levels of Interleukin-6 (IL-6) and its soluble receptors (sIL-6r and sgp130), described in the course of Rheumatoid Arthritis (RA), have been linked to the enhanced activity of this cytokine in this disorder. In this study, the serum concentrations of IL-6 and its soluble receptors were determined in a group of patients with HCV-related arthritis (HCVrA), a condition resembling RA in several aspects, and then compared to those found in a sample of subjects affected by RA. Twenty-one patients with HCVrA, 24 patients with RA and 20 healthy subjects (control group) were examined. Different ELISA methods were used for determination of serum concentrations of IL-6, sIL-6r and sgp130. Increased IL-6 serum levels were found in 15 (71 %) of the patients with HCVrA and in 16 (62 %) of those with RA. Eight (38 %) of the patients with HCVrA and 11 (46%) of those with RA denoted high levels of sIL-6r, while sgp130 levels were elevated in 21 (76%) of the patients with HCVrA and in 16 (69%) of those with RA. A significant difference between the median values of sIL-6r and sgp130 levels in the two groups of patients versus controls was found. A mild correlation of these parameters with RF levels was detected in the RA group. Furthermore, in HCVrA patients the serum levels of IL-6, sIL-6r and sgp130 appeared unrelated to HCV viraemia and to levels of transaminases. The enhanced serum levels of IL-6 in HCVra patients indicate an increased synthesis and hyperactivity of this cytokine in HCVrA, and the substantial similarity of the behaviour of IL-6 and its serum receptors in the two groups of patients suggests common mechanisms with RA, in which the function of I L-6 is central.

Probing the mechanism of incorporation of fluorescently labeled actin into stress fibers.

The mechanism of actin incorporation into and association with stress fibers of 3T3 and WI38 fibroblasts was examined by fluorescent analog cytochemistry, fluorescence recovery after photobleaching (FRAP), image analysis, and immunoelectron microscopy. Microinjected, fluorescein-labeled actin (AF-actin) became associated with stress fibers as early as 5 min post-injection. There was no detectable cellular polarity in the association of AF-actin with pre-existing stress fibers relative to perinuclear or peripheral regions. The rate of incorporation was quantified by image analysis of images generated with a two-dimensional photon counting microchannel plate camera. After equilibration of up to 2 h post-injection, FRAP demonstrated that actin subunits exchanged rapidly between filaments in stress fibers and the surrounding cytoplasm. When co-injected with rhodamine-labeled bovine serum albumin as a control, only actin was detected in the phase-dense stress fibers. The control protein was excluded from fibers and any linear fluorescence of the control was demonstrated as a pathlength artifact. The incorporation of AF-actin into stress fibers was studied by immunoelectron microscopy using anti-fluorescein as the primary antibody and goat anti-rabbit IgG coupled to peroxidase as the secondary antibody. At 5 min post-injection, reaction product was localized periodically in some fibers with a periodicity of approximately 0.75 microns. In large diameter fibers at 5 min post-injection, the analog was seen first on the surface of fibers, with individual filaments resolvable within the core. In the same cell, thinner diameter fibers were labeled uniformly throughout the diameter. By 20 min post-injection, most fibers were uniformly labeled. We conclude that the rate of actin subunit exchange in vivo is extremely rapid with molecular incorporation into actin filaments of stress fibers occurring as early as a few minutes post-injection. Exchange appears to first occur in filaments along the surface of stress fibers and then into more central regions in a periodic manner. We suggest that the periodic localization of actin at very early time points is due to a local microheterogeneity in which microdomains of fast vs. slower incorporation result from the periodic localization of actin-binding protein, such as alpha-actinin, along the length of the fiber.

Distribution of actin in spreading macrophages: a comparative study on living and fixed cells.

The distribution of actin in proteose peptone-elicited murine peritoneal macrophages is examined with fluorescent analog cytochemistry (FAC), immunofluorescence, and electron microscopy (EM). Living adherent macrophages, microinjected with 5- iodoacetamidofluorescence-labeled actin, show a rather uniform distribution of actin with punctuate and linear fluorescence in the thin peripheral areas of the cell. Apparent incorporation of a portion of linear fluorescence in the thin peripheral areas of the cell. Apparent incorporation of a portion of the microinjected actin into the cell's actin cytoskeleton is also demonstrated when microinjected cells are subsequently examined for fluorescein fluorescence after fixation and extraction. However, a substantial perinuclear pool of actin, observed with FAC, is lost when microinjected cells are prepared for immunofluorescence using standard fixation methods. These results suggest that part of the cellular actin, possibly nonfilamentous or oligomeric, can be extracted during the normal preparative steps for immunofluorescence. When the dynamic distributin of actin structures is examined in living cells, extension of the cell's periphery is associated with the formation of punctuate structures. The distribution of the most stable, nonextractable actin structures in fixed cells at different stages of spreading is quantified using rhodamine-labeled phalloidin and antiactin indirect immunofluorescence. At early stages, the rounded cells show cortical bands of fluorescence surrounding the nuclear region with punctuate structures directly above the plane of the attached plasma membrane. At later time periods, fully spread cells contain both punctuate and linear fluorescent structures. Adherent macrophage membranes, a preparation in which the attached membrane and membrane-cortex are isolated by shearing away the unattached plasma membrane and underlying cytoplasm, show punctuate and linear fluorescence when stained with rhodamine-labeled phalloidin. When the same cell remnant is negatively stained and examined with EM, the fluorescent punctuate structures coincide with electron-dense foci and associated radiating thin filaments. We suggest that the optimal approach for elucidating the distribution of cytoskeletal and contractile proteins involved in motile processes is a combined approach using all three techniques. Although each technique is subject to potential artifacts and limitations, the use of FAC can permit the visualization of both the soluble and stabilized components of the cytoskeleton in living, functional cells. A qualitative method for determining differences in local concentrations of proteins is also presented.

Simultaneous application of microbial transglutaminase and high hydrostatic pressure to improve heat induced gelation of pork plasma.

The effects of treating porcine plasma with microbial tranglutaminase (MTGase) under high hydrostatic pressure (HHP) were studied as a means of improving its gel-forming properties when subsequently heated at pH 5.5, near the pH of meats. Plasma containing varying levels of commercial MTGase was pressurized (400MPa, room temperature, pH 7) for different times, and adjusted to pH 5.5 prior to heating to induce gelation. MTGase-treatment under HHP led to greater enhancement of heat-induced plasma gel properties as compared to control samples. The greatest improvements were achieved by pressurising plasma with 43.3U MTGase/g protein for 30min, thereby achieving recoveries of 49% and 63% in fracture force (gel strength) and fracture distance (gel deformability) of the subsequently heat-induced gels, respectively, relative to gel properties obtained by heating untreated plasma at physiological conditions (pH 7.5).

Draft Genome Sequence of Pseudomonas syringae PDD-32b-74, a Model Strain for Ice-Nucleation Studies in the Atmosphere.

We report here the whole genome sequence of Pseudomonas syringae PDD-32b-74, a gammaproteobacterium isolated from cloud water. This microorganism is equipped with ice-nucleation protein and biosurfactant genes that could potentially be involved in physicochemical processes in the atmosphere and clouds.

Draft Genome Sequence of Pseudomonas graminis PDD-13b-3, a Model Strain Isolated from Cloud Water.

The whole genome of Pseudomonas graminis PDD-13b-3, a strain of bacteria isolated from cloud water, was sequenced. This showed that this microorganism is equipped with genes that could potentially be involved in its survival in the atmosphere and clouds: those for oxidative stress and carbon starvation responses, DNA repair, and iron uptake.

Draft Genome Sequence of Rhodococcus enclensis 23b-28, a Model Strain Isolated from Cloud Water.

The whole genome of Rhodococcus enclensis 23b-28, a bacterial strain isolated from cloud water, was sequenced. This microorganism is equipped with genes able to degrade aromatic compounds and could thus play a role in complex organic matter decomposition in cloud water.

Assessment and risk modeling of airborne enteric viruses emitted from wastewater reused for irrigation.

Reclamation of wastewater (WW) for irrigation, after treatment represents a challenge that could alleviate pressure on water resources and address the increasing demand for agriculture. However, the risks to human health must be assessed, particularly those related to human enteric viruses that resist standard treatments in most wastewater treatment plants (WWTP). The risks associated with exposure to viral bioaerosols near WWTP and near agricultural plots irrigated with WW are poorly documented. The objectives of this study were to 1) better characterize human enteric viruses found in bioaerosols near a "standard WWTP" and over fields irrigated with treated WW and 2) propose a numeric model to assess the health risk to populations located close to the irrigated areas, with particular attention to norovirus, which is responsible for most viral gastroenteritis in France. Water and air samples were collected at various locations in the largest French WW-irrigated site near Clermont-Ferrand, at the WWTP entrance and after treatment, in the air above activated sludge basins, and above fields irrigated with WW. Various enteric viruses were found in the water samples collected both before and after treatment. Norovirus was the most abundant with >10e4 genome copies/l (GC/L) before treatment and ~10e3 GC/L after treatment. Low quantities (<10e3GC/m3) were detected in the air above active sludge pools and irrigated plots. Hepatitis E virus was detected in all sampled compartments. A quantitative microbial risk assessment (QMRA) approach, including a simplified atmospheric dispersion model, allowed assessment of norovirus infection risk. The Bayesian QMRA approach considered wind speed measurements over 21years, and the variability and uncertainty of all measurements throughout the chain up to the risk. The probability of infection within one year for the most exposed WWTP employees was >10e-4 for strong wind speed (≥3m/s) and a constant emission rate of 8e3 GC/m3/s. This probability decreases by 3 log when the distance to the emission source is doubled. This information can aid development of safe water reuse policies in terms of local setback distance and wind conditions for wastewater reuse.

Characterization of ice-nucleating bacteria using on-line electron impact ionization aerosol mass spectrometry.

The mass spectral signatures of airborne bacteria were measured and analyzed in cloud simulation experiments at the AIDA (Aerosol Interaction and Dynamics in the Atmosphere) facility. Suspensions of cultured cells in pure water were sprayed into the aerosol and cloud chambers forming an aerosol which consisted of intact cells, cell fragments and residual particles from the agar medium in which the bacteria were cultured. The aerosol particles were analyzed with a high-resolution time-of-flight aerosol mass spectrometer equipped with a newly developed PM2.5 aerodynamic lens. Positive matrix factorization (PMF) using the multilinear engine (ME-2) source apportionment was applied to deconvolve the bacteria and agar mass spectral signatures. The bacteria mass fraction contributed between 75 and 95% depending on the aerosol generation, with the remaining mass attributed to agar. We present mass spectra of Pseudomonas syringae and Pseudomonas fluorescens bacteria typical for ice-nucleation active bacteria in the atmosphere to facilitate the distinction of airborne bacteria from other constituents in ambient aerosol, e.g. by PMF/ME-2 source apportionment analyses. Nitrogen-containing ions were the most salient feature of the bacteria mass spectra, and a combination of C4 H8 N(+) (m/z 70) and C5 H12 N(+) (m/z 86) may be used as marker ions.

Parental divorce and the well-being of children: a meta-analysis.

This meta-analysis involved 92 studies that compared children living in divorced single-parent families with children living in continuously intact families on measures of well-being. Children of divorce scored lower than children in intact families across a variety of outcomes, with the median effect size being .14 of a standard deviation. For some outcomes, methodologically sophisticated studies yielded weaker effect sizes than did other studies. In addition, for some outcomes, more recent studies yielded weaker effect sizes than did studies carried out during earlier decades. Some support was found for theoretical perspectives emphasizing parental absence and economic disadvantage, but the most consistent support was found for a family conflict perspective.

Reversibility bull's-eye: a new polar bull's-eye map to quantify reversibility of stress-induced SPECT thallium-201 myocardial perfusion defects.

Myocardial ischemia is currently interpreted from SPECT thallium-201 (201Tl) tomograms by the subjective visual finding of stress-induced perfusion defects which "normalize" or "reverse" by 4 hr. Thus, we have developed a computer method to quantify and display the three-dimensional distribution of reversible segments. Circumferential profiles generated from the short axis slices are normalized to the reference area in the stress study. The stress is subtracted from the normalized delayed data, and then displayed as a polar bull's-eye plot so that positive values show areas that have "reversed" or "improved." Patient profiles are compared to means and standard deviations of reversibility for all pixels determined from the Emory normal male files. Criteria for reversibility were developed from studies of 42 male patients found to have 48 defects, as determined by the consensus of five blinded expert observers. There was computer agreement with the experts on 25 of 31 relatively fixed and 14 of 17 reversible defects. Our preliminary results indicate that this new method promises to aid observers to more consistently identify and quantify the reversibility of SPECT 201Tl myocardial perfusion defects.

Progesterone induction of calcitonin expression in the murine mammary gland.

Progesterone, via its nuclear receptor, is mandatory not only for the induction and specification of mammary gland ductal side-branching and lobuloalveologenesis but also for carcinogen-induced mammary tumorigenesis. Notwithstanding these recent advances, a more comprehensive molecular explanation of progesterone-induced mammary morphogenesis is contingent upon the identification and characterization of mammary molecular targets that are responsive to the progesterone signal. Toward this goal, we report that calcitonin, a 32 amino acid peptide hormone involved in calcium homeostasis, is exclusively expressed in, and secreted from, luminal epithelial cells within the mammary gland of the pregnant mouse, and, importantly, its expression is progesterone-dependent. Conversely, the calcitonin receptor is present during all stages of post-natal mammary development examined, is localized to the myoepithelial cell lineage, and is not regulated by progesterone. Because calcitonin induction spatiotemporally correlates with increases in progesterone-induced mammary gland proliferation and structural remodeling, we posit that calcitonin - through its receptor - may be involved in one or both of these progesterone-dependent processes.

Effects of high-dose furosemide and small-volume hypertonic saline solution infusion in comparison with a high dose of furosemide as a bolus, in refractory congestive heart failure.

BACKGROUND: Diuretics, have been accepted as first-line treatment in refractory heart failure, but a lack of response is a frequent event. A randomised single blind study was performed to evaluate the effects of the combination of high-dose furosemide and small-volume hypertonic saline solution (HSS) infusion in the treatment of refractory NYHA class IV congestive heart failure (CHF). MATERIALS AND METHODS: Sixty patients (21 F/39 M) with refractory CHF (NYHA class IV) of different etiologies, unresponsive to high oral doses of furosemide, ACE-inhibitors, digitalis, and nitrates, aged 65-90 years, were enrolled. They had to have an ejection fraction (EF) <35%, serum creatinine <2 mg/dl, BUN

Central obesity and hypertension: the role of plasma endothelin.

Hypertension and central obesity are two conditions closely linked, but the mechanisms responsible for obesity-associated hypertension are still unclear. In the last few years, several studies addressed the role of endothelin-1 (ET-1) in the development and maintenance of hypertension. This study was designed to evaluate plasma ET-1 in normotensive and hypertensive central obese subjects compared with a lean healthy group. Our final goal was to analyze the relationship between plasma ET-1, blood pressure, and left ventricular structure and function in central obese subjects (both normotensives and hypertensives). ET-levels have been assessed by the radioimmunoassay method in 20 lean normotensives and in 57 central obese subjects; 30 of them were hypertensives and 27 of them were normotensives. Twenty-four-hour mean blood pressure (MBP/24 h) by noninvasive ambulatory blood pressure monitoring, left ventricular mass/ height (LVM/H), and left ventricular ejection fraction (LVEF) by echocardiography and peak filling rate (PFR) by radionuclide study were also measured. ET levels were significantly (P < .05) higher in obese hypertensives and obese normotensives than in lean normotensives. In addition, ET levels were significantly (P < .05) higher in obese hypertensives than in obese normotensives. ET were directly related to LVM/ H (r = 0.86; P < .001) and MBP/24 h (r = 0.48; P < .009) but only in obese hypertensives. Multiple regression analysis indicated that ET-1 plasma levels remain an independent predictor of MBP/ 24 h and LVM/H also when age was included in the analysis. These data suggest that obesity-associated hypertension is characterized by an endothelial dysfunction that may contribute to the higher cardiovascular risk detectable in these patients.

Effects of chromium picolinate supplementation on insulin sensitivity, serum lipids, and body composition in healthy, nonobese, older men and women.

BACKGROUND: Chromium is an essential nutrient required for carbohydrate and lipid metabolism. Chromium supplementation in humans has been reported to improve glucose metabolism and improve serum lipid parameters and to reduce body fat; parameters that worsen with aging. As a result, chromium picolinate has been widely promoted as a health aid for the general population. The purpose of the study was to examine the effects of chromium supplementation on insulin sensitivity, serum lipids, and body composition in nonobese, healthy men and women of advanced age. METHODS: A randomized, double-blind, placebo-controlled study with 19 subjects (9 men and 10 women), aged 63-77, were given either chromium picolinate, 1,000 microg/d, or a placebo for 8 weeks. Serum lipids were measured at baseline and 8 weeks. Insulin sensitivity and body composition were measured with the minimal-model intravenous glucose tolerance test and dual-energy x-ray absorptiometry scan, respectively, at baseline and after 8 weeks of chromium or placebo supplementation. RESULTS: No significant change in serum lipids, insulin sensitivity, or body composition was observed in the chromium group compared with the placebo group. CONCLUSIONS: Chromium picolinate supplementation alone does not appear to improve insulin sensitivity, serum lipids, or change body composition in nonobese, healthy men and women of advanced age.

Transient ovarian failure: a complication of uterine artery embolization.

OBJECTIVE: To report a case of transient ovarian failure shortly after arterial embolization for treatment of uterine fibroids, followed by recovery of ovarian function. DESIGN: Case report. SETTING: A university-based hospital. PATIENT: A 49-year-old woman with menorrhagia and anemia secondary to uterine fibroids and refractory to medical management. The follicle-stimulating hormone (FSH) level on cycle day 3 before the procedure was 8.2 mIU/mL. INTERVENTION(S): Bilateral uterine artery embolization for treatment of menorrhagia. MAIN OUTCOME MEASURE(S): Serum FSH level. RESULT(S): The patient developed amenorrhea and hot flashes 3 months after uterine artery embolization. Her serum FSH level at that time was 140.1 mIU/mL. Four months later, uterine bleeding resumed; her serum FSH level was 2.1 mIU/mL. CONCLUSION(S): Uterine artery embolization may hasten ovarian failure. This procedure should be reserved for women who have completed their child-bearing or are poor candidates for myomectomy. Patients should be counseled appropriately about the risk of possible ovarian failure.

Predicted values for lung diffusing capacity in healthy children.

Lung diffusing capacity was measured in 92 healthy Caucasian children (52 boys and 40 girls), ranging in age from 6 to 17 years, using the single breath carbon monoxide technique (DLCO). Multiple regression equations were developed to predict the DLCO value. Standing height, gender, weight, and age were significant factors in the prediction of DLCO. Sitting height was not a significant covariate when standing height was in the model, but it could replace standing height in the equation. Equations are presented for the calculation of predicted DLCO in children based on standing or sitting height, age, sex, and weight.

The legacy of parents' marital discord: consequences for children's marital quality.

Drawing on a national longitudinal study of 297 parents and their married offspring, the authors found that parents' marital discord was negatively related to offspring's marital harmony and positively related to offspring's marital discord. The transmission of marital quality was not mediated by parental divorce, life-course variables, socioeconomic attainment, retrospective measures of parent-child relationships, or psychological distress. Offspring's recollections of parental discord, however, mediated about half of the association between parents' reports of marital discord and offspring's reports of discord in their own marriages. Parental behaviors most likely to predict problematic marriages among offspring included jealousy, being domineering, getting angry easily, being critical, being moody, and not talking to the spouse.

Divorce and psychological stress.

While research on adjustment to divorce has been extensive, the paucity of studies assessing stress before and after divorce has kept the relation between psychological stress and martial dissolution unclear. Conflicting findings cast some doubt on the utility of using a crisis model to study divorce. Analysis of three-wave panel data from a national sample of persons married in 1980 indicates that the crisis model is appropriate for understanding adjustment to divorce. Comparisons of divorced persons with married persons show a predivorce rise in stress which then returns to levels comparable to those reported by married individuals. No evidence was found supporting the idea that a high level of psychological stress is a general cause of divorce or that dissolution resulted in more or less permanent elevation of psychological stress. Findings support the hypothesis that predivorce resources and outlooks influence the amount of stress experienced in the two years immediately following divorce. Below median family incomes, no post-high school experience, and wife not in the labor force put divorcing individuals at a disadvantage. Individuals reporting few premarital troubles and beliefs in the immorality of divorce also appear to experience heightened stress in the two years following divorce.

Succinate sumatriptan evaluation by Doppler echocardiography in patients with migraine.

Recent reports show that sumatriptan administration increases blood pressure and vascular resistance both in systemic and pulmonary circulation. This study was performed to evaluate by echo-Doppler technique the hemodynamic effects of subcutaneous sumatriptan administration. Forty-one migraine subjects (26 males, 15 females), mean age 36 +/- 2 years (range 36-39 years), and 20 healthy control subjects (14 males, six females), mean age 36 +/- 2 years (range 36-39 years) were randomized (double-blind) to receiving sumatriptan (group A) or placebo (group B). After a 2-week complete pharmacological washout, clinical examination, electrocardiogram, and Doppler echocardiography were performed at baseline, 15, 30, 45, and 60 min after sumatriptan or placebo administration. No significant differences were found between the two groups regarding Doppler echocardiographic parameters (aortic integral, pulmonary integral, end-systolic and end-diastolic diameters) and heart rate; only a slight but not significant increase in arterial blood pressure was observed in group A. Our data show that succinate sumatriptan can be used with safety in patients without hypertension and other cardiovascular disease.