Prevalence of Pulmonary Infections Caused by Atypical Pathogens in non-HIV Immunocompromised Patients.

Although atypical bacteria are important causes of lower airway infections, data on their role in immunocompromised patients are scarce. The aim of the study was to evaluate the prevalence of atypical pulmonary infections in patients with various types of immunosuppression, and to analyze clinical characteristics of these infections. Eighty non-HIV immunocompromised patients with different underlying diseases and clinical and radiological signs of pulmonary infection were enrolled. Due to incomplete data on eight patients, 72 patients were eligible for final analysis (median age 58 years). All patients underwent fiberoptic bronchoscopy and bronchoalveolar lavage. Bronchoalveolar lavage fluid (BALF) fluid samples were sent for direct microscopy, cultures, and fungal antigen detection, when appropriate. Commercial qualitative amplification assay (PNEUMOTRIS oligomix Alert Kit(®)), based on nested PCR method, was used to detect specific DNA sequences of L. pneumophila, C. pneumoniae, and M. pneumoniae in BALF. There were 61 (84.7 %) patients with hematologic diseases, 3 (4.2 %) after solid organ transplantation, and 8 (11.1 %) with miscellaneous diseases affecting immune status. Specific sequences of M. pneumoniae, C. pneumoniae, and L. pneumophila DNA were found in 7 (9.7 %), 2 (2.8 %), and 0 patients, respectively. In 8 of these patients co-infections with different microorganisms were demonstrated. Co-infection with A. baumanii and P. aeruginosa was diagnosed in three patients who died. We conclude that atypical lower airway infections are uncommon in immunocompromised patients. The majority of these infections are co-infections rather than single pathogen infections.

The effect of opioid agonists and antagonists on gastrointestinal motility in mice selected for high and low swim stress-induced analgesia.

BACKGROUND: The opioid system in the gastrointestinal (GI) tract plays an important physiological role, but is also responsible for the side effect of opioid drugs, including troublesome constipation in chronic pain treatment. The aim of this study was to characterize and validate a new mouse model to study the effects of opioid agonists and antagonists in the GI tract. METHODS: Six-week-old male Swiss-Webster mice, divergently bred for high (HA) and low (LA) swim stress-induced analgesia (SSIA), were used in the study. To assess the influence of opioid agonists (morphine and loperamide) and antagonists (naloxone hydrochloride, NLX and naloxone methiodide, NLXM) on GI motility, whole GI transit (whole GIT) and upper GIT assays were conducted. To evaluate the expression of opioid receptors in the ileum and colon of HA and LA mice, immune staining was performed. KEY RESULTS: The effect of morphine was more pronounced in HA line, whereas loperamide exerted a stronger effect in LA mice. Furthermore, NLX and NLXM differentially abolished the inhibitory action of the central and peripheral opioid system on whole and upper GIT in HA and LA mice. The differences in GI motility between HA and LA mice coexisted with parallel changes in the expression of opioid receptors in the ileum and colon. CONCLUSIONS & INFERENCES: Differences in the activity of the endogenous opioid system are responsible for the vulnerability to changes in GI motility during treatment with opioids. Our findings validate the HA/LA model for further studies on opioids in the GI tract.

Elevated ST2 Distinguishes Incidences of Pediatric Heart and Small Bowel Transplant Rejection.

Elevated serum soluble (s) suppressor of tumorigenicity-2 is observed during cardiovascular and inflammatory bowel diseases. To ascertain whether modulated ST2 levels signify heart (HTx) or small bowel transplant (SBTx) rejection, we quantified sST2 in serially obtained pediatric HTx (n = 41) and SBTx recipient (n = 18) sera. At times of biopsy-diagnosed HTx rejection (cellular and/or antibody-mediated), serum sST2 was elevated compared to rejection-free time points (1714 ± 329 vs. 546.5 ± 141.6 pg/mL; p = 0.0002). SBTx recipients also displayed increased serum sST2 during incidences of rejection (7536 ± 1561 vs. 2662 ± 543.8 pg/mL; p = 0.0347). Receiver operator characteristic (ROC) analysis showed that serum sST2 > 600 pg/mL could discriminate time points of HTx rejection and nonrejection (area under the curve [AUC] = 0.724 ± 0.053; p = 0.0003). ROC analysis of SBTx measures revealed a similar discriminative capacity (AUC = 0.6921 ± 0.0820; p = 0.0349). Quantitative evaluation of both HTx and SBTx biopsies revealed that rejection significantly increased allograft ST2 expression. Pathway and Network Analysis of biopsy data pinpointed ST2 in the dominant pathway modulated by rejection and predicted tumor necrosis factor-α and IL-1ß as upstream activators. In total, our data indicate that alloimmune-associated pro-inflammatory cytokines increase ST2 during rejection. They also demonstrate that routine serum sST2 quantification, potentially combined with other biomarkers, should be investigated further to aid in the noninvasive diagnosis of rejection.

In vitro study of the interaction of heregulin-functionalized magnetic-optical nanorods with MCF7 and MDA-MB-231 cells.

Multifunctional nanoparticles that actively target specific cells are promising tools for cancer diagnosis and therapy. In this article we review the synthesis and surface chemistry of Fe-Au nanorods and their characterization using microscopy. The diameter of the rods used in this study was selected to be 150-200 nm so that they did not enter the cells. The 80 nm-long Au tips of the nanorods were functionalized with heregulin (HRG), and the micron-long Fe portion was coated with a poly(ethylene glycol) monolayer to minimize non-specific interactions. Nanorods functionalized with HRG were found to preferentially bind to MCF7 cells that express high levels of the receptor tyrosine-protein kinase ErbB2/3. Magnetic tweezers measurements were used to characterize the kinetic properties of the bond between the HRG on the rods and ErbB2/3 on the surface of the cells. The strong magnetization of Fe-Au nanorods makes them excellent candidates for in-vitro and in-vivo imaging, and magnetic therapeutic applications targeting cancer cells in circulation.

Digital transplantation pathology: combining whole slide imaging, multiplex staining and automated image analysis.

Conventional histopathology is the gold standard for allograft monitoring, but its value proposition is increasingly questioned. "-Omics" analysis of tissues, peripheral blood and fluids and targeted serologic studies provide mechanistic insights into allograft injury not currently provided by conventional histology. Microscopic biopsy analysis, however, provides valuable and unique information: (a) spatial-temporal relationships; (b) rare events/cells; (c) complex structural context; and (d) integration into a "systems" model. Nevertheless, except for immunostaining, no transformative advancements have "modernized" routine microscopy in over 100 years. Pathologists now team with hardware and software engineers to exploit remarkable developments in digital imaging, nanoparticle multiplex staining, and computational image analysis software to bridge the traditional histology-global "-omic" analyses gap. Included are side-by-side comparisons, objective biopsy finding quantification, multiplexing, automated image analysis, and electronic data and resource sharing. Current utilization for teaching, quality assurance, conferencing, consultations, research and clinical trials is evolving toward implementation for low-volume, high-complexity clinical services like transplantation pathology. Cost, complexities of implementation, fluid/evolving standards, and unsettled medical/legal and regulatory issues remain as challenges. Regardless, challenges will be overcome and these technologies will enable transplant pathologists to increase information extraction from tissue specimens and contribute to cross-platform biomarker discovery for improved outcomes.

[The diagnosis of disorders of the T-cell immunity component in space flight]. / Diagnostika narushenii T-kletochnogo zvena immuniteta v kosmicheskom polete.

The investigation yielded two modifications of the procedure FOR determination of T-lymphocytes' functional activity in microsamples of the finger capillary blood and PHA-assisted cultivation of cells in syringe for subsequent morphological analysis by cosmonauts in flight. Testing of the proposed modifications in 14 healthy donors with unconstrained work/rest schedule, 4 volunteers in 60- and 6 volunteers in 120-day HDT showed a rather high comparability of data obtained with the modified and genuine procedures for determination of the T-lymphocytes proliferative activity in cosmonauts before and after orbital flights. Thus, the modified procedures can be successfully used for monitoring the immune status of cosmonauts on missions, and individuals exposed to extreme environments.

[Human cellular immunity and space flights]. / Kletochnyi immunitet cheloveka i kosmicheskii polet.

Results of studying cellular immunity of crew members of long-term space missions, data of an experiment with extended head-down tilting of human subjects (HDT), and data obtained in the course of adapting immunologic test methods for the use in microgravity are summarized. Disorders in immunologic reactivity were shown to occur under the conditions of space flight. They included decreases in both quantitative and functional indices of cellular immunity, and emergence of signs of sensibilization to different allergens. The modified tests were evaluated by efficacy of determination of the proliferative activity of lymphocytes in minimal volumes of capillary blood inside one-piece syringes with a medium containing PHA and the Cytodex-1 suspension, evaluation of natural cytotoxicity on the level of an effector cell (number of peripheral blood lymphocytes capable of producing conjugates with fixed target cells). Sensibilization to allergens of normal human microflora was tested in analogs of routine hematocrit capillaries used for examination of healthy donors under ordinary rest-work regimen. No significant differences between the standard and modified tests were revealed. The proposed test modifications are quite simple in use and require minimum of equipment.

Clinical and echocardiographical study of the aortic homograft implantations in patients with Marfan syndrome.

The aim of the study was to assess the long-term results of surgical treatment with homogenic aortic grafts (HAGs) implantation in patients with Marfan syndrome. There were 31 patients with Marfan syndrome and aortic aneurysm who were operated on between 1980 and 1996. Aortic dissection was diagnosed in 14 patients, DeBakey Type I in six patients and Type II in eight patients. Four patients had to be operated urgently in cardiogenic shock with cardiac tamponade. Sealing up and reinforcement with strip of felt or Gore-Tex has been applied in 22 patients. The surgical modifications mentioned above have been applied since 1987 in all patients with the diameter of the aortic ring exceeding 30 mm or with active infective endocarditis or during reoperation. In 16 patients the space between the aortic homograft and patients own aortic wall was joined to the right atrial auricle. Patients were followed up for 12-179 months (average: 94.6 +/- 499). Three patients died in the early postoperative period and four patients died in the late postoperative period. Rethoracotomy because of bleeding complications was necessary in five patients. HAG damage was responsible for six other reoperations-new HAGs have been implanted in three patients and artificial prostheses were implanted in the other three patients. In the late follow-up period significant improvement in cardiac performance was observed in 24 patients (NYHA I or II). Survival probability of 15 years for the whole group was 80%. The lowest survival probability has been shown in the group of patients with DeBakey Type I aortic dissection (35% survived 15 years after operation). Echocardiographic follow-up has shown that the pressure gradient in HAG was low (7.4 +/- 6.2 mmHg). Only in two patients did the HAG gradient exceeded 20 mmHg. There were no significant differences concerning aortic ring diameters, dimensions of HAG and echocardiographic parameters between the group with surgical modifications, i.e. sealing up and reinforcement with strip of felt or Gore-Tex applied and the group in which these modifications were not applied. Homogenic aortic graft implantation as a method of surgical treatment of aortic aneurysm in patients with Marfan syndrome avoids postoperative anticoagulation, results in substantial improvement of cardiac performance and prolongs life. Surgical treatment should be considered in asymptomatic patients with large aneurysms (exceeding 55-65 mm) in patients with Marfan syndrome because there is a high risk of death in this group of patients in the case of dissection.

[Immune resistance of humans during extended isolation]. / Immunologicheskaia rezistentnost' cheloveka pri dlitel'noi izoliatsii.

Analyzed were parameters of the immune reactivity and allergological status of participants in the 135-day chamber experiment with periods of induced psychoemotional strain. Stressful situations gave rise to shifts in the antibody system, i.e. decreased blood levels of ImgA, ImgM, and ImgG, decline in the functional activity of T-lymphocytes, suppression of the cytokinetic activity of lymphocytes-natural killers, and sensibilization to various allergens. Totality of these shifts resembles the ones which are typically seen during initial readaptation to the Earth's conditions on after long-term space mission.

Successful full-term pregnancy in a patient three and a half years after a heart transplant.

The patient is a 28 year old woman who received a heart transplant in 1992 secondary to hypertrophic cardiomyopathy with unremarkable post-operative course. In the period immediately post transplantation the patient was on a four-drug immunosuppressive regimen which was subsequently changed to standard three-agent therapy. This therapy was continued until the patient became pregnant. In the first trimester only Cyclosporine (CsA) was used, and thereafter, the patient was continued on the previous three agent regimen. Toward the end of pregnancy a rise in systolic pressure was observed, but the child was delivered by spontaneous vaginal delivery without complications in the 38th week of pregnancy. The newborn weighed 3320 g and was in good health. A sharp fall in the newborn CsA blood levels was observed post delivery reaching zero level on the third day of life. At the present time, both mother and baby are in good health, 6 weeks after delivery.

[Infectious aortic valve endocarditis. Echocardiographic evaluation]. / Infekcyjne zapalenie wsierdzia zastawki aortalnej. Ocena echokardiograficzna.

In the group of 37 patients (pts) with infective endocarditis of aortic valve comparative analysis of echocardiograms with intraoperative or pathomorphological findings was performed. Infection caused development of vegetations in 17 pts, cusp rupture in 7 and perivalvular abscess in 4 pts. Echocardiographic examination enabled diagnosis of vegetations in 16 pts. In 2 pts abscess cavity was shown between mitral and aortic annulus. Two-dimensional echocardiography provided more detailed data concerning number and localization of vegetation, and development of perivalvular abscess cavity. However cusps rupture was shown by M-mode echograms in 4 from 7 pts. In the course of antibiotic therapy 7 pts died: from 30 pts treated surgically the result of treatment was beneficial in 27. The examinations confirmed poor clinical prognosis of premature mitral valve closure for the patients with aortic insufficiency.

[Immunologic mechanisms of regulating calcium metabolism in human bone tissue (production of osteoclast-activating factor) during long-term anti-orthostatic hypokinesia]. / Immunologicheskie mekhanizmy reguliatsii obmena kal'tsiia v kostnoi tkani cheloveka (produktsiia osteoklastaktiviruiushchego faktora) pri dlitel'noi antiortostaticheskoi gipokinezii.

During 370 days 10 healthy male subjects were exposed to antiorthostatic hypokinesia. The exposure enhanced the production of the osteoclast-activating factor (OAF) by blood mononuclear cells. The enhancement began after the 2nd month of hypokinesia and continued to increase thereafter. The index of resorption was: prior to hypokinesia--1.15 +/- 0.156, 8 months after onset--2.05 +/- 0.129, and 11.5 months after onset--2.37 +/- 0.296, the upper normal limit being 1.54. The changes were correlated with a greater amount of active T-lymphocytes in blood and a progressive increase of calcium negative balance. This investigation lends support to the previously formulated hypothesis (I. V. Konstantinova, 1986) that immunological mechanisms associated with the OAF production contribute to calcium metabolism disorders in human bone during prolonged space flights.

Immunocompetent cells producing humoral mediators of bone tissue mineral metabolism during space flight simulation.

During 120-day and 370 day hypokinesia the possibility has been shown of manifestation of mechanism of immunological regulation of osteoclast functions. Supernatants of mononuclear peripheral blood cells which were in vitro nitrogen-stimulated, had an increased potential for resorption in 45Ca-labelled mice fetus long bone organ cultures. Resorbing activity was increased in mononuclear supernatants from some of the subjects exposed to 120-day hypokinesia and from all subjects exposed to 370-day hypokinesia. This variable returned to base-line values after completion of the bed-rest period. Lymphocyte in vitro proliferative activity decreased at the end of hypokinesia and during the initial days of recovery, and the number of active T-rosetting cells was, on the contrary increased. This suggested a possible activation of part of immunocompetent cell population potentially producing humoral regulators of bone cell functions in vivo. A study of a group of healthy males performing their routine daily activity and of patients with local osteoporosis (paradontitis) showed significant differences between normal subjects and patients, confirming the informative value of the method used and allowed to establish approximate limits of physiological variations of the values.

Efficiency of nodule initiation in cowpea and soybean.

When serial dilutions of a suspension of Bradyrhizobium japonicum strain 138 were inoculated onto both soybean and cowpea roots, the formation of nodules in the initially susceptible region of the roots of both hosts was found to be linearly dependent on the log of the inoculum dosage until an optimum dosage was reached. Approximately 30- to 100-fold higher dosages were required to elicit half-maximal nodulation on cowpea than on soybean in the initially susceptible zone of the root. However, at optimal dosages, about six times as many nodules formed in this region on cowpea roots than on soybean roots. There was no appreciable difference in the apparent rate of nodule initiation on these two hosts nor in the number of inoculum bacteria in contact with the root. These results are consistent with the possibility that cowpea roots have a substantially higher threshold of response to symbiotic signals from the bacteria than do soybean roots. Storage of B. japonicum cells in distilled water for several weeks did not affect their viability or efficiency of nodule initiation on soybean. However, the nodulation efficiency of these same cells on cowpea diminished markedly over a 2 week period. These differential effects of water storage indicate that at least some aspects of signal production by the bacteria during nodule initiation are different on the two hosts. Mutants of B. japonicum 138 defective in synthesis of soybean lectin binding polysaccharide were defective in their efficiency of nodule initiation on soybean but not on cowpea. These results also suggest that B. japonicum may produce different substances to initiate nodules on these two hosts.

A solid-phase radio-binding assay for the characterization of lectin recognition.

A rapid and quantitative method for characterizing lectin specificity is presented. This assay is analogous to solid-phase radioimmune assays and utilizes the property of irreversible binding of proteins to vinyl microtiter plates. The lectins phytohemagglutinin and concanavalin A bind to vinyl and retain their biological properties. Iodinated fetuin and immunoglobulin G are both bound by the immobilized lectins. Competing amounts of ligands presented at the same time as the iodinated glycoproteins are shown to reduce binding to the immobilized lectins. Conversely, glycoprotein ligands immobilized to vinyl retain their ability to be recognized and bind lectins. The solid-phase binding assay can be used to characterize the ligand-binding specificity of lectins. For example, the pattern of glycoprotein inhibition of the binding of iodinated gorgonian lectin from Leptogorgia virgulata to insolubilized thyroglobulin is virtually identical to the pattern reported previously using hemagglutination inhibition. The solid-phase radio-binding assay is rapid, reproducible, and sensitive to nanogram quantities of ligand added. Most importantly, it provides quantitative information on lectin recognition.

[Effect of space flight on cosmonaut peripheral blood lymphocyte blast transformation]. / Vliianie kosmicheskogo poleta na reaktsiiu blasttransformatsii limfotsitov perifericheskoi krovi kosmonavtov.

The postflight investigation of cell mediated immunity of seven cosmonauts showed significant increase in spontaneous activity of lymphocytes of peripheral blood and decrease in stimulation of lymphocytes cultured with PHA at R + O. Variations in these parameters were transient, remaining within physiological limits. Two cosmonauts displayed a stable increase in lymphocyte activity during blast transformation test with Staphylococcus aureus culture filtrate.