SESSION TYPE: Infectious Disease Global Case Report PostersPRESENTED ON: Tuesday, October 23, 2012 at 01:30 PM - 02:30 PM

INTRODUCTION:

The chest radiological features of Mycoplasma pneumoniae (M. pneumonia) pneumonia are varied. The characteristics of the host cell-mediated immunity (CMI) might influence the pattern of pulmonary lesions in M. pneumoniae infection#1

CASE PRESENTATION:

A previously healthy nonsmoking 40-year-old woman with two-day history of fever and productive cough was admitted to a local hospital. Two weeks before admission, her daughter had M. pneumoniae infection with mild but persistent symptoms despite of macrolide treatment. The patient was initially treated as pneumonia with a combination of azithromycin 500mg q 24hr for 3 days and IV ceftriaxone 2g q24hr followed by sulbactam/ampicillin 3g q6hr for one week. However, her clinical symptoms, desaturation with persistent high fever and chest radiological findings were refractory to the treatment and she was referred to our hospital. On examination, there were markedly diminished breath sounds over the left lung and scattered crackle on the right lower lung. Soluble Interleulin-2 receptor (s-IL2R) was remarkably elevated, and tuberculin skin test was negative. The High-resolution computed tomography (HRCT) of the chest revealed airspace consolidation and atelectasis in the left lung and mild centrilobular nodules in the right lower lung. Immediately administration of methylprednisolone 1g for three days was initiated with azithromycin, 500mg q 24hr and IV Ciprofloxacin (CPFX), 300mg q12hr. Although these resulted in a dramatic improvement of the clinical symptoms and airspace consolidation, diffuse centrilobular nodules in the left lung with wheezing remained. Gram's stain, routine bacterial and fungal cultures were negative. The acute serum cold agglutinin titer was 1:16384 and the M pneumoniae IgG (complement fixation) was 1:64.and IgM (particle agglutination) was 1: 20480. The sputum polymerase chain reaction for determination of M pneumoniae was positive. We concluded that she had exclusive M pneumoniae pneumonia. Finally, the patient was treated with azithromycin for 14days and IV CPFX for 7days with prednisone, 40mg for 7days. Six weeks after the treatment, wheezing and chest radiological manifestation in the left lung still remained. Pulmonary function test indicated peripheral airway obstruction without airway reversibility. Moreover, HRCT on expiration revealed the mosaic pattern, indicating air trapping. These results suggested the bronchiolitis obliterans (BO), as the sequeale of M. pneumoniae pneumonia, although transbronchial lung biopsy could not elucidated the histology of BO. Bronchoalveolar lavage fluid revealed infiltration of neutrophil and lymphocyte.

DISCUSSION:

Herein we describe an adult with exclusive M pneumoniae pneumonia who presented with the dramatic change of the radiological manifestations, T cell marker activation, negative tuberculin test, suggesting the possibility of the cell-mediated immunity (CMI) of the host. Her clinical course was very different from that of her daughter. In general, the chest CT findings can be divided into the two groups: one group had a predominance of nodular opacities with a centrilobular distribution and the other showed a predominance of an airspace consolidation. Our case showed airspace consolidation in the early stage, changed nodular opacities in the late stage, and demonstrated remarkable elevation of s-IL2R and reduction to the treatment, as a T cell activation marker. Negative tuberculin test might suggest the lung local CMI decline. All these findings were influenced by the treatment with corticosteroids and macrolide antibiotics, and host CMI. Corticosteroids had been successfully used for both radiological types of M. pneumoniae infection for the purpose of anti-inflammatory effects, without no strong evidence. Macrolide antibiotics also had been used not only for direct antimicrobial activity but also anti-inflammatory effects. These treatment might lead to a successful outcome in our case.

CONCLUSIONS:

Radiological manifestations of M. pneumoniae infection could be influenced by both the treatment and the host CMI response.1) Eur Respir J, 1996, 9, 669-672DISCLOSURE: The following authors have nothing to disclose: Hideki Makino, Haruki Kobayashi, Kei Nakashima, Nobuhiro Asai, Naoko Katsurada, Masafumi Misawa, Norihiro Kaneko, Masahiro AoshimaNo Product/Research Disclosure InformationKameda Medical Center, Kamogawa, Japan.

The display of glucose oxidase (GOx) on yeast cell surface using a-agglutinin as anchor motif was successfully developed. Both the immunochemical analysis and enzymatic assay showed that active GOx was efficiently expressed and translocated on the cell surface. Compared with conventional GOx, the yeast cell surface displayed GOx (GOx-yeast) demonstrated excellent enzyme properties such as good stability within a wide pH range (pH3.5-11.5), good thermostability (retaining over 94.8% enzyme activity at 52 oC and 84.2% enzyme activity at 56 oC), and high D-glucose specificity. Additionally, direct electrochemistry was achieved at GOx-yeast/multiwalled-carbon-nanotube modified electrode, suggesting the host cell of yeast did not have any adverse effect on the electrocatalytic property of the recombinant GOx. Thus, a novel electrochemical glucose biosensor based on this GOx-yeast was developed. The as-prepared biosensor was linear with the concentration of D-glucose within the range of 0.1-14 mM and a low detection limit of 0.05 mM (S/N=3). Moreover, the as-prepared biosensor is stable, specific, reproducible, simple, and cost-effective, which can be applicable for real samples detection. The proposed strategy to construct robust GOx-yeast may be applicable to explore other oxidase-displaying system based whole-cell biocatalysts, which can find broad potential application in biosensor, bioenergy and industrial catalysis.

The modular synthesis of seven libraries containing 51 self-assembling amphiphilic Janus dendrimers with the monosaccharides D-mannose, D-galactose and the disaccharide D-lactose in their hydrophilic part is reported. These unprecedented sugar-containing dendrimers are named amphiphilic Janus glycodendrimers. Their self-assembly by simple injection of THF or ethanol solution into water or buffer and by hydration was analyzed by a combination of methods including dynamic light scattering, confocal microscopy, cryogenic transmission electron microscopy, Fourier transform analysis, and micropipette-aspiration experiments to assess mechanical properties. These libraries revealed a diversity of hard and soft assemblies, including unilamellar spherical, polygonal and tubular vesicles denoted glycodendrimersomes, aggregates of Janus glycodendrimers and rod-like micelles, named glycodendrimer aggregates and glycodendrimermicelles, cubosomes denoted glycodendrimercubosomes, as well as solid lamellae. These assemblies are stable over time in water and in buffer, exhibit narrow molecular-weight distribution and display dimensions that are programmable by the concentration of the solution they are injected from. This study elaborated the molecular principles leading to single-type soft glycodendrimersomes assembled from amphiphilic Janus glycodendrimers. The multivalency of glycodendrimersomes with different size and their ligand bioactivity were demonstrated by selective agglutination with a diversity of sugar-binding protein receptors such as the plant lectins concanavalin A and the highly toxic mistletoe Viscum album L. agglutinin, the bacterial lectin PA-IL from Pseudomonas aeruginosa, and, of special biomedical relevance, human adhesion/growth-regulatory galectin-3 and galectin-4. These results demonstrated the candidacy of glycodendrimersomes as new mimics of biological membranes with programmable glycan ligand presentations, as supramolecular lectin blockers, vaccines, and targeted delivery devices.

Lycoris is aurea agglutinin (LAA) has attracted rising attention due to its remarkable bioactivities. Here, we aimed at investigating its anti-tumor activities.In vitro methods including MTT, cellular morphology observation, FCM and immunoblotting were performed. In vivo methods like detection of tumor volume, body weight and survival ratio, as well as TUNEL staining were performed.LAA triggers G2 /M phase cell cycle arrest via up-regulating p21expression as well as down-regulating cdk-1cyclinA singling pathway, and induces apoptotic cell death through inhibiting PI3K-Akt survival pathway in human lung adenocarcinoma A549 cells. While LAA has no significant cytotoxic effect toward normal human embryonic lung fibroblast HELF cells, and moreover, LAA could amplify the antineoplastic effects of cisplatin toward A549 cells. Lastly LAA also bears anti-cancer and apoptosis-inducing effects in vivo, and it could decrease the volume and weight of subcutaneous tumor mass obviously as well as expand lifespan of mice. These findings may provide a new perspective for elucidating the complicated molecular mechanisms of LAA-induced cancer cell growth-inhibition and death, providing a new opportunity of LAA as a potential candidate anti-neoplastic drug for future cancer therapeutics.

Autoimmune hemolytic anemia (AIHA) is an infrequent group of diseases defined by autoantibody mediated red blood cell destruction. Correct diagnosis and classification of this condition are essential to provide appropriate treatment. AIHA is divided into warm and cold types according to the characteristics of the autoantibody involved and by the presence of an underlying or associated disorder into primary and secondary AIHA. Due to its low frequency, treatment for AIHA is largely based on small prospective trials, case series, and empirical observations. This review describes in detail the different treatment approaches for autoimmune hemolytic anemia. Warm antibody type AIHA should be treated with steroids, to which most patients respond, although relapse can occur and maintenance doses are frequently required. Splenectomy is an effective second line treatment and can provide long-term remission without medication. Rituximab is a useful alternative for steroid refractory patients, those requiring high maintenance doses and unfavorable candidates for surgery. Promising therapeutic modifications with this monoclonal antibody are emerging including drug combinations, lower doses, and long-term use. Primary cold agglutinin disease has been recognized as having a lymphoproliferative monoclonal origin. It is unresponsive to both steroids and splenectomy. Rituximab is currently the best therapeutic alternative for this condition, and several treatment regimens are available with variable responses.

The mechanisms by which aldosterone increases Na(+), K(+) ATPase and sodium channel activity in cortical collecting duct and distal nephron have been extensively studied. Recent investigations demonstrate that aldosterone increases Na-H exchanger-3 (NHE-3) activity, bicarbonate transport, and H(+) ATPase in proximal tubules. However, the role of aldosterone in regulation of Na(+), K(+) ATPase in proximal tubules is unknown. We hypothesize that aldosterone increases Na(+), K(+) ATPase activity in proximal tubules through activation of the mineralocorticoid receptor (MR). Immunohistochemistry of kidney sections from human, rat, and mouse kidneys revealed that the MR is expressed in the cytosol of tubules staining positively for lotus tetragonolobus agglutinin and type IIa sodium-phosphate cotransporter (NpT2a), confirming proximal tubule localization. Adrenalectomy in Sprague Dawley rats decreased expression of MR, ENaC α, Na(+), K(+) ATPase α1, and NHE-1 in all tubules, while supplementation with aldosterone restored expression of above proteins. In human kidney proximal tubule (HKC11) cells, treatment with aldosterone resulted in translocation of MR to the nucleus and phosphorylation of SGK-1. Treatment with aldosterone also increased Na(+), K(+) ATPase-mediated (86)Rb uptake and expression of Na(+), K(+) ATPase α1 subunits in HKC11 cells. The effects of aldosterone on Na(+), K(+) ATPase-mediated (86)Rb uptake were prevented by spironolactone, a competitive inhibitor of aldosterone for the MR, and partially by mifepristone, a glucocorticoid receptor (GR) inhibitor. These results suggest that aldosterone regulates Na(+), K(+) ATPase in renal proximal tubule cells through an MR-dependent mechanism.

The binding of the stromal cell-derived factor-1α (SDF-1α) to the cysteine (C)-X-C motif chemokine receptor 4 (CXCR4) has emerged as a key signal for stem and progenitor cells trafficking to the circulation from the bone marrow. Our aim was to investigate the role of daily intermittent administration of AMD3100 (a specific reversible CXCR4 receptor antagonist) during the healing process after myocardial infarction (MI). Wistar rats were subjected to MI and AMD3100 was injected intraperitoneally after surgery. SDF-1α mRNA expression was measured by real-time polymerase chain reaction. Histology changes were analyzed with immunofluorescence, Masson's trichrome staining, and wheat germ agglutinin. The number of leukocytes in peripheral blood was measured by complete blood cell count analysis. The activities of matrix metalloproteinase-2/9 (MMP-2/9) were determined by gelatin zymography. The expression level of SDF-1α mRNA in the infarcted tissue was enhanced rapidly (6 h), peaked at 24 h, and then declined to the normal level at 7 days post-MI. AMD3100 further enhanced the increase of SDF-1α in infarct area. Increased leukocytes were observed in AMD3100-treated groups. The mobilization of c-kit(+) stem/progenitor cells and enhanced neovascularization were augmented by AMD3100. Additionally, AMD3100 improved ventricular remodeling, which was revealed by the decrease of infarct size, viable cardiomyocyte cross-sectional area and left ventricle (LV) expansion index, and the increase of LV free wall thickness. The activities of MMP-2/9 were up-regulated by AMD3100. In conclusion, short-term intermittent administration of AMD3100 could accelerate the wound healing process in experimental MI and be a potential therapy for the treatment of MI.

The urinary bladder is innervated by parasympathetic preganglionic neurons (PPNs) that express μ-opioid receptors (MOR) in the sacral parasympathetic nucleus (SPN) at lumbosacral segments L6-S1. The SPN also contains endomorphin 2 (EM2)-immunoreactive (IR) fibers and terminals. EM2 is the endogenous ligand of MOR. In the present study, retrograde tract-tracing with cholera toxin subunit b (CTb) or wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) via the pelvic nerve combined with immunohistochemical staining for EM2 and MOR to identify PPNs within the SPN as well as synaptic connections between the EM2-IR terminals and MOR-expressing PPNs in the SPN of the rat. After CTb was injected into the pelvic nerve, CTb retrogradely labeled neurons were almost exclusively located in the lateral part of the intermediolateral gray matter at L6-S1 of the lumbosacral spinal cord. All of the them also expressed MOR. EM2-IR terminals formed symmetric synapses with MOR-IR, WGA-HRP-labeled and WGA-HRP/MOR double-labeled neuronal cell bodies and dendrites within the SPN. These results provided morphological evidence that EM2-containing axon terminals formed symmetric synapses with MOR-expressing PPNs in the SPN. The present results also show that EM2 and MOR might be involved in both the homeostatic control and information transmission of micturition.

The primary production site of erythropoietin (EPO) is shifted from the liver to the kidney shortly after birth. Under conditions of lost or reduced kidney production, it is valuable to measure the production capacity of the liver. However, there is a lack of urine or serum based methods that can distinguish endogenous EPO produced in different cell types. Here is presented a method based on chromatographic interaction with the lectin wheat germ agglutinin (WGA) that can distinguish presumably liver-produced EPO, found in anaemic patients receiving epoetin and darbepoetin, from kidney-produced EPO found in healthy individuals. All the tested samples from haemodialysis patients with end-stage renal disease showed a presence of liver EPO. In some samples, the liver-produced EPO made up 90-100% of total EPO at a concentration of 8-10ng/L in urine, which indicates that the liver has a quite high production capacity, although not adequate for the degree of anaemia. This glycoform analysis has made it possible to affirm that some anaemic patients can increase their liver-production of EPO. The use of such a method can give better insight into the regulation of non-renal endogenous EPO production, a potential source of EPO intended to replace administration of exogenous EPO.

Mucins and mucin-type glycoproteins, collectively referred to as mucin-type O-glycans, are implicated in many important biological functions and pathological conditions, including malignancy. Presently, there is no reliable method to measure the total mucin-type O-glycans of a sample, which may contain one or more of these macromolecules of unknown structures. We report the development of an improved microassay that is based on the binding of lectins to the unique and constant GalNAc-Ser/Thr structural feature of mucin-type O-glycans. Since the sugar-amino acid linkage in the mucin-type O-glycans is invariably cryptic, we first chemically removed the heterogeneous peripheral and core saccharides of model glycoconjugates before examining for their interactions using an enzyme-linked lectin assay (ELLA). Desialylation of the model glycoconjugates led to maximal binding of the lectins but additional treatments such as Smith degradation did not result in increased binding. Of the lectins tested for their ability to probe the desialylated O-glycans, jacalin showed the highest sensitivity followed by champedak galactose binding (CGB) lectin and Vicia villosa agglutinin. Further improvement in the sensitivity of ELLA was achieved by using microtiter plates that were pre-coated with the CGB lectin, which increased the specificity of the assay to mucin-type O-glycans. Finally, the applicability of the developed sandwich ELLA to crude samples was demonstrated by estimating trace quantities of the mucin-type O-glycans in the human serum.