Variations in the Red Cells in Paroxysmal Nocturnal Haemoglobinuria

S ummary . The red cells of 22 patients with paroxysmal nocturnal haemoglobinuria have been evaluated by the complement lysis sensitivity test of Rosse & Dacie. Four patterns of abnormal susceptibility to complement lysis were seen. In Group 1 (10 patients), markedly abnormal complement-sensitive cells were present and cells of a second population were not distinguishable from normal cells. In Group 2 (four patients), markedly abnormal complement-sensitive cells were present and the second population of cells was clearly distinguishable, but not markedly different from normal. In Group 3 (five patients), three populations of cells were present: a markedly abnormal complement-sensitive population, a population of moderately abnormal cells and a population of normal-appearing cells. In Group 4 (three patients), no cells with marked complement sensitivity were seen; two populations were present, one was apparently normal and the other moderately abnormal (3–4 times as sensitive as normal). During the period of observation two patients in Group 4 acquired a markedly abnormal complement-sensitive population.
The proportion of cells in each population varied during the course of the illness. In three patients the proportion of complement-sensitive cells increased during the onset of the illness, and in two patients the proportion decreased gradually during the resolution of the disease. Increase in the total production of red cells resulted in an increased proportion of complement-sensitive cells, as did decrease in haemolytic rate. Increase in the haemolytic rate resulted in a decreased proportion of complement-sensitive cells.     

Ischemic-appearing electrocardiographic changes predict myocardial injury in patients with intracerebral hemorrhage

Objectives

Myocardial injury is common among patients with intracerebral hemorrhage (ICH). However, it is challenging for emergency physicians to recognize acute myocardial injury in this population, as electrocardiographic (ECG) abnormalities are common in this setting. Our objective is to examine whether ischemic-appearing ECG changes predict subsequent myocardial injury in the context of ICH.

Methods

Consecutive patients with primary ICH presenting to a single academic center were prospectively enrolled. Electrocardiograms were retrospectively reviewed by 3 independent readers. Anatomical areas of ischemia were defined as I and aVL; II, III, and aVF; V₁ to V₄; and V₅ and V₆. Medical record review identified myocardial injury, defined as troponin I or T elevation (cutoff 1.5 and 0.1 ng/mL, respectively), within 30 days.

Results

Between 1998 and 2004, 218 patients presented directly to our emergency department and did not have a do-not-resuscitate/do-not-intubate order; arrival ECGs and troponin levels were available for 206 patients. Ischemic-appearing changes were noted in 41% of patients, and myocardial injury was noted in 12% of patients. Ischemic-appearing changes were more common in patients with subsequent injury (64% vs 37%; P = .02). After multivariable analysis controlling for age and cardiac risk factors, ischemic-appearing ECG changes independently predicted myocardial injury (odds ratio, 3.2; 95% confidence interval, 1.3-8.2). In an exploratory analysis, ischemic-appearing ECG changes in leads I and aVL as well as V₅ and V₆ were more specific for myocardial injury (P = .002 and P = .03, respectively).

Conclusion

In conclusion, although a range of ECG abnormalities can occur after ICH, the finding of ischemic-appearing changes in an anatomical distribution can help predict which patients are having true myocardial injury.     

Surgery for retroperitoneal soft tissue sarcomas: aggressive re-resection of recurrent disease is possible

INTRODUCTION

Retroperitoneal soft tissue sarcomas represent a relatively rare and complex therapeutic problem where surgery forms the mainstay of treatment and is technically demanding. In this study, we review a single UK centre’s experience with the surgical management of retro-peritoneal soft tissue sarcoma.

PATIENTS AND METHODS

We present analysis of data on patients treated between 1997 and 2006, our first 75 patients. Data collected from the Access database, included patient demographics, staging modalities, peri-operative details, treatment, outcome, pathological diagnosis and subsequent complications.

RESULTS

A total of 75 patients (M:F, 44:31) underwent 115 resectional procedures as part of the management of retroperitoneal soft-tissue sarcoma. There were 12 major complications for the 115 procedures (morbidity of 8.69%). The 30-day operative mortality was zero and the 90-day mortality rate was 1.33% (1/75). Follow-up ranged from 16–131 months. The median disease-free survival was 69 months (range, 59-78 months). Recurrences developed in 46 patients; median time to overall recurrence was 13 months (range, 3-71 months). Of these 46, 22 developed localised recurrence, which was amenable to further resection. In the cohort of patients with recurrent disease, median survival in those who underwent surgery was 53 months (range, 30–76 months) and median survival in those who did not undergo surgery was 30 months (range, 18–41 months) and this difference was statistically significant (log rank, P = 0.01).

CONCLUSIONS

Extensive resectional surgery with minimal morbidity, devoid of mortality is feasible in the treatment of retroperitoneal sarcoma. Development of recurrent disease is a significant factor influencing survival; however, localised recurrences are amenable to surgery and this can lead to improved survival.     

Protection Efficacy of Multivalent Egg Yolk Immunoglobulin against Eimeria tenella Infection in Chickens

Background

To control avian coccidiosis with drug-independent strategy effectively and safely, multivalent hyperimmune egg yolk immunoglobulin (IgY) was prepared and its ability to protect against Eimeria tenella infection was evaluated.

Methods

Hens were orally immunized with live oocysts of 5 species of Eimeria for six times, antibody titers in serum and yolk were monitored by indirect enzyme-linked immunosorbent assay. The specific IgY was isolated, purified and lyophilized. IgY powder was orally administrated as dietary supplement in newly hatched chicks at various dosages. Birds were orally challenged with 10000 sporulated oocysts of E. tenella at 10 days of age, weighed and killed at 8 days post challenge, and the protective effect was assessed.

Results

The averge yeid of IgY was 9.2 mg/ml yolk, the antibody titer of IgY reached to 1:163840 per mg with the purity up to 98%. Chickens fed IgY resulted in reduced mortality, increased body weight gain (BWG), reduced oocyst shedding, reduced caecal lesion score and increased anti-coccidial index. In terms of BWG and caecal lesion, IgY significantly enhanced the resistance of bird at ≥ 0.05% of IgY in the diet when compared with the challenged control group (P<0.05). No significant difference was observed at dosage ≥ 0.5% and 1.0% when BWG and caecal lesion were compared with the sodium salinomycin control group, respectively (P>0.05).

Conclusion

Supplementing newly hatched chicks with Eimeria-specific IgY represents a promising strategy to prevent avian coccidiosis.     

Tumor suppressor Lzap regulates cell cycle progression,doming, and zebrafish epiboly

Initial stages of embryonic development rely on rapid, synchronized cell divisions of the fertilized egg followed by a set of morphogenetic movements collectively called epiboly and gastrulation. Lzap is a putative tumor suppressor whose expression is lost in 30% of head and neck squamous cell carcinomas. Lzap activities include regulation of cell cycle progression and response to therapeutic agents. Here, we explore developmental roles of the lzap gene during zebrafish morphogenesis. Lzap is highly conserved among vertebrates and is maternally deposited. Expression is initially ubiquitous during gastrulation, and later becomes more prominent in the pharyngeal arches, digestive tract, and brain. Antisense morpholino‐mediated depletion of Lzap resulted in delayed cell divisions and apoptosis during blastomere formation, resulting in fewer, larger cells. Cell cycle analysis suggested that Lzap loss in early embryonic cells resulted in a G2/M arrest. Furthermore, the Lzap‐deficient embryos failed to initiate epiboly—the earliest morphogenetic movement in animal development—which has been shown to be dependent on cell adhesion and migration of epithelial sheets. Our results strongly implicate Lzap in regulation of cell cycle progression, adhesion and migratory activity of epithelial cell sheets during early development. These functions provide further insight into Lzap activity that may contribute not only to development, but also to tumor formation. Developmental Dynamics 240:1613–1625, 2011. © 2011 Wiley‐Liss, Inc.     

Activation of PAR2 receptors sensitizes primary afferents and causes leukocyte rolling and adherence in the rat knee joint

Background and Purpose

The PAR₂ receptors are involved in chronic arthritis by mechanisms that are as yet unclear. Here, we examined PAR₂ activation in the rat knee joint.

Experimental Approach

PAR₂ in rat knee joint dorsal root ganglia (DRG) cells at L3-L5, retrogradely labelled with Fluoro-gold (FG) were demonstrated immunohistochemically. Electrophysiological recordings from knee joint nerve fibres in urethane anaesthetized Wistar rats assessed the effects of stimulating joint PAR₂ with its activating peptide, 2-furoyl-LIGRLO-NH₂ (1–100 nmol·100 μL⁻¹, via close intra-arterial injection). Fibre firing rate was recorded during joint rotations before and 15 min after administration of PAR₂ activating peptide or control peptide. Leukocyte kinetics in the synovial vasculature upon PAR₂ activation were followed by intravital microscopy for 60 min after perfusion of 2-furoyl-LIGRLO-NH₂ or control peptide. Roles for transient receptor potential vanilloid-1 (TRPV1) or neurokinin-1 (NK₁) receptors in the PAR₂ responses were assessed using the selective antagonists, SB366791 and RP67580 respectively.

Key Results

PAR₂ were expressed in 59 ± 5% of FG-positive DRG cells; 100 nmol 2-furoyl-LIGRLO-NH₂ increased joint fibre firing rate during normal and noxious rotation, maximal at 3 min (normal; 110 ± 43%, noxious; 90 ± 31%). 2-Furoyl-LIGRLO-NH₂ also significantly increased leukocyte rolling and adhesion over 60 min. All these effects were blocked by pre-treatment with SB366791 and RP67580 (P < 0.05 compared with 2-furoyl-LIGRLO-NH₂ alone).

Conclusions and Implications

PAR₂ receptors play an acute inflammatory role in the knee joint via TRPV1- and NK₁-dependent mechanisms involving both PAR₂-mediated neuronal sensitization and leukocyte trafficking.     

Importance of Antibodies to Lipopolysaccharide in Natural and Vaccine-Induced Serum Bactericidal Activity against Neisseria meningitidis Group B

Analysis of the specificity of bactericidal antibodies in normal, convalescent, and postvaccination human sera is important in understanding human immunity to meningococcal infections and can aid in the design of an effective group B vaccine. A collection of human sera, including group C and group B convalescent-phase sera, normal sera with naturally occurring cross-reactive bactericidal activity, and some postvaccination sera, was analyzed to determine the specificity of cross-reactive bactericidal antibodies. Analysis of human sera using a bactericidal antibody depletion assay demonstrated that a significant portion of the bactericidal activity could be removed by purified lipopolysaccharide (LPS). LPS homologous to that expressed on the bactericidal test strain was most effective, but partial depletion by heterologous LPS suggested the presence of antibodies with various degrees of cross-reactivity. Binding of anti-L3,7 LPS bactericidal antibodies was affected by modification of the core structure, suggesting that these functional antibodies recognized epitopes consisting of both core structures and lacto-N-neotetraose (LNnT). When the target strain was grown with 5′-cytidinemonophospho-N-acetylneuraminic acid (CMP-NANA) to increase LPS sialylation, convalescent-phase serum bactericidal titers were decreased by only 2- to 4-fold, and most remaining bactericidal activity was still depleted by LPS. Highly sialylated LPS was ineffective in depleting bactericidal antibodies. We conclude that natural infections caused by strains expressing L3,7 LPS induce persistent, protective bactericidal antibodies and appear to be directed against nonsialylated bacterial epitopes. Additionally, subsets of these bactericidal antibodies are cross-reactive, binding to several different LPS immunotypes, which is a useful characteristic for an effective group B meningococcal vaccine antigen.