Inflammatory Fluid Research Articles

Fulminant Tuberculous Meningitis

A 56-year-old Japanese male presented with tuberculous meningitis (TBM) manifesting as irrational behavior. He underwent lumbar puncture, which showed inflammatory cerebrospinal fluid (CSF) findings. Administration of anti-tuberculosis (TB) agents was started on the day after admission (Day 1) because delayed treatment of TBM might be fatal. On Day 4, magnetic resonance (MR) imaging and MR angiography showed fresh infarctions, hydrocephalus, and stenoses of arteries. CSF drainage and biopsy of brain tissue were performed, but the pathological findings were non-specific. Frequent CSF examinations, cultures, and polymerase chain reaction were performed, but no positive finding of TB was obtained. He died on Day 14. Brain autopsy showed Langerhans giant cells and Ziehl-Neelsen-positive TB bodies. Unfortunately, our patient suffered very poor outcome irrespective of early anti-TB treatment starting Day 1, suggesting the probability of delayed admission or drug-resistant TB. TB infection including TBM has become rare in developed countries, and diagnosis remains difficult. Corticosteroid therapy may be effective for TBM, but may be restricted and ameliorate mortality but not morbidity. Further study is required to establish second line treatment if TBM is resistant to anti-TB agents and corticosteroid administration.

Defining Autoimmune Epilepsy

The goal of this study was to identify clinical characteristics and response to treatment of autoimmune epilepsy. The authors retrospectively reviewed data from 32 patients followed in a specialty autoimmune neurology clinic. The authors define autoimmune epilepsy as (1) epilepsy as the exclusive or predominant symptom, and (2) a suspected autoimmune pathogenesis based on detection of neural autoantibodies, inflammatory cerebrospinal fluid (CSF) findings (leukocytosis or …

Inflammatory synovial fluid microenvironment drives primary human chondrocytes to actively take part in inflammatory joint diseases

The role of human chondrocytes in the pathogenesis of cartilage degradation in rheumatic joint diseases has presently gained increasing interest. An active chondrocyte participation in local inflammation may play a role in the initiation and progression of inflammatory joint diseases and in a disruption of cartilage repair mechanisms resulting in cartilage degradation. In the present study, we hypothesized that inflammatory synovial fluid triggers human chondrocytes to actively take part in inflammatory processes in rheumatic joint diseases. Primary human chondrocytes were incubated in synovial fluids gained from patients with rheumatoid arthritis, psoriasis arthritis and reactive arthritis. The detection of vital cell numbers was determined by using Casy Cell Counter System. Apoptosis was measured by Annexin-V and 7AAD staining. Cytokine and chemokine secretion was determined by a multiplex suspension array. Detection of vital cells showed a highly significant decrease in chondrocyte numbers. Flow cytometry demonstrated a significant increase in apoptotic chondrocytes after the incubation. An active secretion of cytokines such as MCP-1 and MIF by chondrocytes was observed. The inflammatory synovial fluid microenvironment mediates apoptosis and cell death of chondrocytes. Moreover, in terms of cytokine secretion, it also induces an active participation of chondrocytes in ongoing inflammation.

Membrane-associated CD93 regulates leukocyte migration and C1q-hemolytic activity during murine peritonitis.

CD93 is emerging as a novel regulator of inflammation; however, its molecular function is unknown. CD93 exists as a membrane-associated glycoprotein on the surface of cells involved in the inflammatory cascade, including endothelial and myeloid cells. A soluble form (sCD93) is detectable in blood and is elevated with inflammation. In this study, we demonstrate heightened susceptibility to thioglycollate-induced peritonitis in CD93(-/-) mice. CD93(-/-) mice showed a 1.6-1.8-fold increase in leukocyte infiltration during thioglycollate-induced peritonitis between 3 and 24 h that returned to wild type levels by 96 h. Impaired vascular integrity in CD93(-/-) mice during peritonitis was demonstrated using fluorescence multiphoton intravital microscopy; however, no differences in cytokine or chemokine levels were detected with Luminex Multiplex or ELISA analysis. C1q-hemolytic activity in CD93(-/-) mice was decreased by 22% at time zero and by 46% 3 h after thioglycollate injection, suggesting a defect in the classical complement pathway. Leukocyte recruitment and C1q-hemolytic activity was restored to wild type levels when CD93 was expressed on either hematopoietic cells or nonhematopoietic cells in bone marrow chimeric mice. However, elevated levels of sCD93 in inflammatory fluid were observed only when CD93 was expressed on nonhematopoietic cells. Because cell-associated CD93 was sufficient to restore a normal inflammatory response, these data suggest that cell-associated CD93, and not sCD93, regulates leukocyte recruitment and complement activation during murine peritonitis.

Heparin affects the interaction of kininogen on endothelial cells

In the plasma kallikrein-kinin system, it has been shown that when plasma prekallikrein (PK) and high molecular weight kininogen (HK) assemble on endothelial cells, plasma kallikrein (huPK) becomes available to cleave HK, releasing bradykinin, a potent mediator of the inflammatory response. Because the formation of soluble glycosaminoglycans occurs concomitantly during the inflammatory processes, the effect of these polysaccharides on the interaction of HK on the cell surface or extracellular matrix (ECM) of two endothelial cell lines (ECV304 and RAEC) was investigated. In the presence of Zn+2, HK binding to the surface or ECM of RAEC was abolished by heparin; reduced by heparan sulfate, keratan sulfate, chondroitin 4-sulfate or dermatan sulfate; and not affected by chondroitin 6-sulfate. By contrast, only heparin reduced HK binding to the ECV304 cell surface or ECM. Using heparin-correlated molecules such as low molecular weight dextran sulfate, low molecular weight heparin and N-desulfated heparin, we suggest that these effects were mainly dependent on the charge density and on the N-sulfated glucosamine present in heparin. Surprisingly, PK binding to cell- or ECM-bound-HK and PK activation was not modified by heparin. However, the hydrolysis of HK by huPK, releasing BK in the fluid phase, was augmented by this glycosaminoglycan in the presence of Zn2+. Thus, a functional dichotomy exists in which soluble glycosaminoglycans may possibly either increase or decrease the formation of BK. In conclusion, glycosaminoglycans that accumulated in inflammatory fluids or used as a therapeutic drug (e.g., heparin) could act as pro- or anti-inflammatory mediators depending on different factors within the cell environment.

Encéphalite à anticorps anti-récepteur du NMDA : une entité traitable impliquant les lymphocytes B. Deux observations

Introduction N-methyl-D-aspartate receptor antibody (anti-NMDA-r AB) encephalitis has been recently identified. We report two cases illustrating the clinical features, response to immunomodulatory treatment and involvement of B-lymphocytes that characterizes this disorder. Case reports These patients illustrated the classic clinical features of anti-NMDA-r AB encephalitis including occurrence in young female, presence of severe neurological and psychiatric manifestations with confusion, seizures, mutism, hypovigilence and involuntary movements, and inflammatory cerebrospinal fluid. Both patients improved after immunotherapy. In case 1, the encephalitis was associated with an ovarian teratoma containing neuronal elements. In case 2, there was no tumor identified. A brain biopsy showed prominent perivascular B-cells infiltrates with some T-cells distributed in the brain parenchyma. Conclusion Anti-NMDA-r AB encephalitis is certainly not rare and needs to be promptly recognized and treated. An associated neoplasia is inconstant and the pathophysiology involves humoral immunity.

Serum regulates cortisol bioactivity by corticosteroid‐binding globulin‐dependent and independent mechanisms, as revealed by combined bioassay and physicochemical assay approaches

Corticosteroid-binding globulin (CBG) is the principal carrier of natural glucocorticoids in the circulation, and we hypothesized that it modulates glucocorticoid bioactivity (GBA). Alterations in CBG, the presence of noncortisol, naturally occurring glucocorticoids and the use of potent, synthetic glucocorticoids, all make it difficult to assess adrenal activity in-vivo; these problems can be addressed by a glucocorticoid bioassay. A bioassay was developed for serum GBA and a physicochemical ultrafiltration-liquid chromatography-tandem mass spectrometry assay for free serum cortisol (FreeF). We studied individuals homozygous and heterozygous for a nonfunctioning CBG variant (CBG G237V) and healthy controls. FreeF concentrations were similar in healthy controls, and those with absent functional CBG, but surprisingly we found low GBA in CBG null individuals. This may suggest that CBG delivers cortisol to target cells. However, further experiments revealed that dilution of serum in the bioassay caused release of cortisol from CBG, resulting in elevated GBA measurements in all but the CBG G237V homozygotes. Furthermore, we identified a specific and potent inhibitory effect of high concentration serum on glucocorticoid sensitivity of the recipient cells used in the bioassay. Analysis of inflammatory synovial fluid, a filtrate of serum with lower CBG concentration, revealed elevated free cortisol compared to noninflammatory synovial fluid, a change not attributable to interconversion between cortisol and cortisone. Our findings reveal that dilution of CBG enhances cortisol release, and so bioactivity, and also that serum potently induces glucocorticoid resistance in target cells.

Degenerative and inflammatory markers in the cerebrospinal fluid of multiple sclerosis patients with relapsing-remitting course of disease and after clinical isolated syndrome

Background: Literature includes evidence of initial predominance of inflammation and later development of neurodegeneration in the pathogenesis of multiple sclerosis (MS).Objective: To search for differences in inflammatory and degenerative cerebrospinal fluid (CSF) markers between relapsing-remitting MS (RRMS) and the clinical isolated syndrome (CIS).Methods: A total of 148 subjects were evaluated, 65 MS patients (45 RRMS and 20 CIS) and 83 controls. The evaluated parameters included albumin quotient and prealbumin, transferrin, C3 and C4 complement factors, haptoglobin, beta-2-microglobulin, orosomucoid, alpha-1-antitrypsin, apolipoprotein A-I, apolipoprotein B, cystatin C, neuron-specific enolase, tau protein, beta-amyloid, oligoclonal IgG band (OCB), and IgG quotient (QIgG).Results: No differences were found in the inflammatory and degenerative CSF markers between patients with RRMS and CIS. QIgG was higher in RRMS than that in CIS but the number of OCB was higher after CIS. Cystatin C levels were significantly lower in RRMS compared to the other groups. It can be considered an indicator of the demyelination degree. Normal values of beta-amyloid were less frequent in RRMS compared to those in controls.

Efficacy of Topically Delivered Moxifloxacin against Wound Infection by Pseudomonas aeruginosa and Methicillin-Resistant Staphylococcus aureus

Wound infection is a common risk for patients with chronic nonhealing wounds, causing high morbidity and mortality. Currently, systemic antibiotic treatment is the therapy of choice, despite often leading to several side effects and the risk of an insufficient tissue penetration due to impaired blood supply. If systemically delivered, moxifloxacin penetrates well into inflammatory blister fluid, muscle, and subcutaneous adipose tissues and might therefore be a possible option for the topical treatment of skin and infected skin wounds. In this study, topical application of moxifloxacin was investigated in comparison to mupirocin, linezolid, and gentamicin using a porcine wound infection and a rat burn infection model. Both animal models were performed either by an inoculation with methicillin-resistant Staphylococcus aureus (MRSA) or Pseudomonas aeruginosa. Wound fluid, tissue, and blood samples were taken, and bacterial counts as well as the moxifloxacin concentration were determined for a 14-day follow-up. A histological comparison of the rat burn wound tissues was performed. Both strains were susceptible to moxifloxacin and gentamicin, whereas mupirocin and linezolid were effective only against MRSA. All antibiotics showed efficient reduction of bacterial counts, and except with MRSA, infected burn wounds reached bacterial counts below 10(5) CFU/g tissue. Additionally, moxifloxacin was observed to promote wound healing as determined by histologic analysis, while no induction of bacterial resistance was observed during the treatment period. The use of topical antibiotics for the treatment of infected wounds confers many benefits. Moxifloxacin is therefore an ideal candidate, due to its broad antibacterial spectrum, its high efficiency, and its potential to promote wound healing.

Contrast visibility for indirect MR arthrography with different protein contents and agent relaxivities at different field strengths: An in vitro model

Protein binding and relaxivity are major determinants of the relative effectiveness of an MR arthrographic contrast agent. We sought to evaluate the optimal concentrations of high and usual relaxivity agents in two different proteinous environments at variable field strength for two MR contrast agents of different relaxivities. At 1.5, 3.0 and 7.0 T, gadobenate dimeglumine (Multihance) with high-relaxivity in proteinous environment and gadoteridol (Prohance) with more typical behavior were studied at 1.25, 2.5, 5, and 10 mmol in 1.7 g/dL and 3g/dL albumin (mimicking protein content of normal and inflammatory synovial fluids, respectively) vs. pure normal saline, as a control. Analysis of image signal intensity (SI) and relaxivity values was done. In our study a change in concentration had no significant effect on T1 SI. In contrast, nearly every change in concentration led to a significant change in T2 SI. In 1.25 mmol concentration, there was no effect on T1 SI of either protein concentrations while higher concentrations showed significant decreased SI in either protein carrier compared to saline. The SI of Gadoteridol was significantly higher (p<0.0001) than that of gadobenate at each of 3T and 7 T, but was significantly lower (p<0.001) at 1.5 T in saline solution while this was not significant for either protein carrier. Both protein carriers had significant effect on T1 (p=0.0124) and T2 (p=0.0118) relaxivities. Also solution concentration significantly (p<0.01) affected both T1 and T2 relaxivities. Field strength did not affect T1 relaxivity (p=0.02511) while it significantly affected T2 relaxivity (p<0.001). This was significant (p=0.035) in case of gadoteridol at 3T. 1.25 mmol concentration of both gadoteridol and gadobenate solutions yields the best diagnostic T1 SI specially in higher fields (3T and 7 T) and avoid the deleterious effect of increasing concentration on T2 SI. Gadoteridol is suggested on 3T field indirect MR arthrograms. Protein had no positive effect on either SI or relaxivities in any joint model.

Retracted] The Beneficial Effect of Direct Peritoneal Resuscitation on Septic Shock in Rats

The high mortality associated with conventionally resuscitated septic shock and the subsequent multiple-organ failure remain a very significant and costly clinical problem. Conventional simple intravenous resuscitation (CR) from septic shock often fails to restore the progressive splanchnic vasoconstriction and hypoperfusion, and fails to reverse gut-derived systemic inflammatory response and fluid sequestration. Numerous interventions have been used to protect organ systems and cellular viability from the lethal injury accompanying hypoperfusion and ischemia but none of these efforts have been sufficient to halt or reverse the main course of the pathophysiology noted with conventional resuscitated shock. Recently, some studies have found that in hemorrhagic shock, direct peritoneal resuscitation (DPR) not only produces sustained hyperperfusion in viscera but also has immunomodulatory and anti-fluid sequestration effects. Although the etiology and pathogenesis of septic shock and hemorrhagic shock differ, both kinds of shock result in hypoperfusion of the intestines and other internal organs. In this paper, we seek to determine whether DPR has a similar therapeutic effect on septic shock/resuscitation.

Inflamed vermiform appendix within the sac of incarcerated left inguinal hernia

A 54-year-old male complained of a continuous pain together with an irreducible swelling of the left inguinal region 8 hours prior to admission to the surgical emergency department. His physical examination revealed a very painful, erythematous, irreducible swelling in the left inguinal region without abdominal peritoneal irritation. Routine blood tests disclosed mild leukocytosis. Abdominal plain X-ray film was not specific, and ultrasonography revealed a 10 cm in length inactive, edematous intestinal section within the inguinal hernia. With the diagnosis of strangulated inguinal hernia, he underwent surgical exploration through a transverse inguinal incision. By opening the hernia sac, 6-8 cc inflammatory fluid drained out, and an inflamed vermiform appendix adhered to the inner surface of the sac was seen. Appendicectomy and primary hernia repair were performed at the same time through the inguinal incision. The postoperative course was uneventful, and the histological examination of the specimen revealed an inflamed appendix.

The K1 Capsular Polysaccharide of Acinetobacter baumannii Strain 307-0294 Is a Major Virulence Factor

Acinetobacter baumannii is a pathogen of increasing medical importance with a propensity to be multidrug resistant, thereby making treatment challenging. Little is known of virulence traits in A. baumannii. To identify virulence factors and potential drug targets, random transposon (Tn) mutants derived from the A. baumannii strain AB307-0294 were screened to identify genes essential for growth in human ascites fluid in vitro, an inflammatory exudative fluid. These studies led to the identification of two genes that were predicted to be required for capsule polymerization and assembly. The first, ptk, encodes a putative protein tyrosine kinase (PTK), and the second, epsA, encodes a putative polysaccharide export outer membrane protein (EpsA). Monoclonal antibodies used in flow cytometric and Western analyses confirmed that these genes are required for a capsule-positive phenotype. A capsule-positive phenotype significantly optimized growth in human ascites fluid, survival in human serum, and survival in a rat soft tissue infection model. Importantly, the clearance of the capsule-minus mutants AB307.30 (ptk mutant, capsule minus) and AB307.45 (epsA mutant, capsule minus) was complete and durable. These data demonstrated that the K1 capsule from AB307-0294 was an important protectin. Further, these data suggested that conserved proteins, which contribute to the capsule-positive phenotype, are potential antivirulence drug targets. Therefore, the results from this study have important biologic and translational implications and, to the best of our knowledge, are the first to address the role of capsule in the pathogenesis of A. baumannii infection.

Intra‐articular opioid analgesia is effective in reducing pain and inflammation in an equine LPS induced synovitis model

Intra-articular administration of morphine as a local analgesic and anti-inflammatory drug is widely used in human medicine. In equids, little is known about its clinical analgesic and anti-inflammatory efficacy. To use an inflammatory orthopaedic pain model to investigate the analgesic and anti-inflammatory effects of intra-articularly administered morphine as a new treatment modality in horses with acute arthritis. In a crossover study design, synovitis was induced in the left or right talocrural joint by means of intra-articular injection of 0.5 ng lipopolyssacharide (LPS). The effect of 120 mg morphine, intra-articularly administered at 1 h after induction of synovitis, was evaluated using both physiological and behavioural pain variables. Synovial fluid was sampled at 0, 4, 8, 28 and 52 h after induction of synovitis and analysed for total protein concentration, leucocyte count and for prostaglandin E(2), bradykinin and substance P concentrations by ELISA. Ranges of motion of metatarsophalangeal and talocrural joints were measured as kinematic variables with the horses walking and trotting on a treadmill under sound and lame conditions. Clinical lameness scores and several behavioural variables related to the perception of pain were obtained. LPS injection caused marked transient synovitis, resulting in increased concentrations of inflammatory synovial fluid markers, clinical lameness, joint effusion and several behavioural changes, such as increased time spent recumbent, decreased limb loading at rest and decreased time spent eating silage. Intra-articular morphine resulted in a significant decrease in synovial white blood cell count, prostaglandin E(2) and bradykinin levels and improvement in clinical lameness, kinematic and behavioural parameters, compared to placebo treatment. Intra-articular morphine offers potent analgesic and anti-inflammatory effects in horses suffering from acute synovitis. Local administration of opioids may be useful for horses with acute inflammatory joint pain and offers possibilities for multimodal analgesic therapies without opioid-related systemic side effects.

Abstract 2939: Leukocyte chemoattractant chemerin as a novel immunotherapeutic agent

Abstract Cancer immunotherapy offers the potential of potent anti-tumor responses while avoiding non-specific toxicities associated with traditional cytotoxic chemotherapy. Localization of key immune cells to tumor sites has been shown to be important for anti-tumor efficacy. Chemerin is a recently described chemoattractant initially isolated from human inflammatory fluids. Chemokine-like receptor 1 (CMKLR1) constitutes the main cellular receptor for chemerin, and is expressed by macrophages, natural killer (NK) cells, and immature dendritic cells (DCs), but has not been shown on T lymphocytes. Chemerin is synthesized and circulates systemically as an inactive precursor, prochemerin, but can be rapidly converted into its active form by proteolytic cleavage of carboxy-terminal residues by multiple proteases. To evaluate chemerin as a potential immunotherapy, we performed intratumoral injections of recombinant active murine chemerin that resulted in significantly decreased tumor growth in a B16 melanoma as well as JC breast tumor models. Tumors lines showed no expression of CMKLR1 by flow cytometric (FACS) analysis, and treatment with exogenous recombinant murine chemerin had no effect on in vitro proliferation. Compared to control media, B16 tumor-conditioned serum-free media was able to attract CMKLR1-transfected L1.2 cells (L1.2/CMKLR1) after the addition of inactive, recombinant murine prochemerin, indicating the ability of tumor-derived factors to activate prochemerin. Treatment with the pan-matrix metalloproteinase (MMP) inhibitor GM6001 resulted in reduced chemotaxis, suggesting the activation of chemerin was in part mediated by MMPs. This supports the ability of tumors to activate prochemerin. B16, as well as additional mouse tumor lines (JC, CT26, EMT6), were then transfected to constitutively express chemerin. Experiments using all tumor lines showed consistent and significantly decreased growth of chemerin-expressing tumors compared with control tumors. There were greater numbers of leukocytes in both the tumors and draining lymph nodes compared to controls. CMKLR1 knockout mice we generated showed abrogated response to chemerin-expressing tumors, supporting the hypothesis that the anti-tumor effects are mediated by this receptor. CD4 and CD8 T lymphocytes from tumor-bearing mice were isolated and shown, for the first time, to have significant CMKLR1 expression compared to control lymphocytes from naïve mice. Incubation of naïve T cells with tumor-conditioned media recapitulated these results, suggesting CMKLR1 expression can be induced by tumor-derived factors. These data suggest not only that tumor-derived proteases can activate chemerin, but also that chemerin expression at tumor sites results in the recruitment of CMKLR1-positive cells, potentially including CMKLR1+ lymphocytes, and establishment of an effective anti-tumor immune response. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2939.

Isotopic labeling of terminal amines in complex samples identifies protein N-termini and protease cleavage products

Effective proteome-wide strategies that distinguish the N-termini of proteins from the N-termini of their protease cleavage products would accelerate identification of the substrates of proteases with broad or unknown specificity. Our approach, named terminal amine isotopic labeling of substrates (TAILS), addresses this challenge by using dendritic polyglycerol aldehyde polymers that remove tryptic and C-terminal peptides. We analyze unbound naturally acetylated, cyclized or labeled N-termini from proteins and their protease cleavage products by tandem mass spectrometry, and use peptide isotope quantification to discriminate between the substrates of the protease of interest and the products of background proteolysis. We identify 731 acetylated and 132 cyclized N-termini, and 288 matrix metalloproteinase (MMP)-2 cleavage sites in mouse fibroblast secretomes. We further demonstrate the potential of our strategy to link proteases with defined biological pathways in complex samples by analyzing mouse inflammatory bronchoalveolar fluid and showing that expression of the poorly defined breast cancer protease MMP-11 in MCF-7 human breast cancer cells cleaves both endoplasmin and the immunomodulator and apoptosis inducer galectin-1.

The Role of Endoscopy in the Management of Cystic Pancreatic Lesions

Pancreatic cystic lesions may be due to pseudocysts and related inflammatory fluid collections, simple cysts, cystic tumours such as serous cystadenoma, mucinous cystic neoplasm and intraductal papillary mucinous neoplasm, as well as solid tumours with areas of cystic degeneration. The main diagnostic challenge is to distinguish premalignant and malignant cystic lesions from benign cystic lesions. Cross-sectional imaging using computer tomography and magnetic resonance imaging/ magnetic resonance cholangiopancreatography provides the initial morphological characterization. Endoscopic ultrasound (EUS) is an important tool when diagnostic doubts persist and is crucial in the assessment of invasive malignancy. EUS-guided fine needle aspiration and cyst fluid analysis has been shown to be cost-effective for risk stratifying the malignant potential of cystic tumours and the need for surgical resection. In the management of symptomatic pseudocysts and related fluid collections, endoscopic drainage has been established as the preferred technique, with efficacy similar to surgery but lower costs and morbidity.

Development and validation of a high performance liquid chromatography method to determine linezolid concentrations in pig pulmonary tissue

Linezolid is the first synthetic compound of a new group of antimicrobials, the oxazolidinones, which inhibit protein synthesis. It shows a broad spectrum of activity against Gram positive organisms. With respect to its pharmacokinetics, linezolid shows a relatively high volume of distribution and good penetration into inflammatory fluids, bone, fat and muscle. A reversed-phase isocratic high-performance liquid chromatographic method for linezolid analysis in piglet pulmonary tissue is described. Tissue samples and controls were prepared in 1 x TBE (1 M Tris, 0.9 M boric acid, 0.01 M EDTA). The mobile phase consisted of 20% ultrafiltered water and 80% of (A) 15 mM potassium monohydrogen phosphate buffer (pH = 5) with (B) acetonitrile (80%/20%; v/v). Samples were homogenized and precipitated with HClO(4) 3% (1/1, v/v). The injection volume was 100 microL. Ofloxacin was used as an internal standard. The assay was linear over a linezolid concentration range: 1.6-100 microg/mL. The method provided good validation data (n = 15): inaccuracy (3.6%), intra and inter-day variability (4.2% and 5.2%, respectively), recovery (91.8%), limit of detection (0.8 microg/mL) and quantitation (1.6 microg/mL) and acceptable stability within 24 h in the auto-sampler. The method offers a fast and simple approach to determine linezolid in pulmonary tissue which could be of use in pharmacokinetic studies.

Severe Cysticercal Meningitis: Clinical and Imaging Characteristics

In disease-endemic areas, severe cysticercal meningitis (SCM) is characterized by intense inflammatory cerebrospinal fluid (CSF) and negative bacterial and fungal cultures. There have been no systematic studies of SCM. We characterized patients with SCM and compare them with neurocysticercosis (NC) patients with mild CSF abnormalities by conducting a nine-year retrospective review at a neurological referral center. Two groups of patients were compared: group A, those with severe CSF pleocytosis > 1,000 cells/mm(3) (n = 12), and group B, those with CSF pleocytosis <or= 1,000 cells/mm(3) (n = 126). All patients had positive CSF results in an enzyme-linked immunosorbent assay for cysticercal antigens and negative CSF cultures for bacteria, fungi, and mycobacteria. Intracranial hypertension, meningeal signs, CSF hypoglycorrachia, and a longer clinical course of NC were more frequently seen in group A. It is likely that SCM often goes unrecognized. Its correct identification may reduce morbidity and risks of unnecessary surgery in patients with chronic NC and CSF shunts.

Methicillin-Resistant Staphylococcus Aureus And Chronic Sinusitis

The increasing incidence of methicillin-resistant Staphylococcus aureus (MRSA) in patients with acute and chronic sinusitis is becoming a clinical concern. Not much is known about the role of MRSA in sinonasal infection and how best to manage it. We describe two patients who developed MRSA sinusitis and failed prolonged treatment with trimethoprim/sulfamethoxazole. Following treatment that involved intravenous daptomycin, both patients fully recovered. Daptomycin is a lipopeptide antibacterial agent with good activity against many Gram-positive pathogens, including MRSA. In addition, daptomycin penetrates inflammatory blister fluid moderately well.