The role of the complement system in innate immunity

Complement is a major component of innate immune system involved in defending against all the foreign pathogens through complement fragments that participate in opsonization, chemotaxis, and activation of leukocytes and through cytolysis by C5b-9 membrane attack complex. Bacterias and viruses have adapted in various ways to escape the complement activation, and they take advantage of the complement system by using the host complement receptors to infect various cells. Complement activation also participates in clearance of apoptotic cells and immune, complexes. Moreover at sublytic dose, C5b-9 was shown to promote cell survival. Recently it was also recognized that complement plays a key role in adaptive immunity by modulating and modifying the T cell responses. All these data suggest that complement activation constitutes a critical link between the innate and acquired immune responses.     

The role of complement activation in atherosclerosis

Atherosclerosis is a chronic inflammatory disease in which dyslipidemia, inflammation, and the immune system play an important pathogenetic role. A role in atherogenesis was demonstrated for monocyte/macrophages, complement system, and T-lymphocytes. Complement activation and C5b-9 deposition occurs both in human and experimental atherosclerosis. Complement C6 deficiency has a protective effect on diet-induced atherosclerosis, indicating that C5b-9 assembly is required for the progression of atherosclerotic lesions. The maturation of atherosclerotic lesions beyond the foam cell stage was shown to be strongly dependent on an intact complement system. C5b-9 may be responsible for cell lysis, and sublytic assembly of C5b-9 induces smooth muscle cell (SMC) and endothelial cell (EC) activation and proliferation. All these data suggest that activation of the complement system plays an important role in atherogenesis.     

A novel mitochondria-targeting method using special staining for the detection of apoptotic hepatocytes

ABSTRACT

Early detection of apoptotic cells on histological slides is of major importance for both diagnostic and research areas. In the current study, the aim was to propose a convenient method to stain the mitochondria and establish whether hepatocytes undergoing apoptosis can be identified in tissue sections using the proposed method. Liver tissue from five adult chinchillas was fixed with 10% neutral buffered formalin for Goldner’s trichrome (GT) and Groat’s iron hematoxylin and eosin (HE) stains and with Kolster’s fixative for the Heidenhain’s iron hematoxylin procedure. The HE and GT-stained sections showed the morphological features consistent with apoptosis i.e., homogenous intensely acidophilic cytoplasm, cell shrinkage with an irregular outline, nuclear shrinkage with cloudy karyoplasm, and karyopyknosis in the late stage. Sections stained with Heidenhain’s iron hematoxylin method was used to pinpoint mitochondria and revealed cells which were undergoing the first stages of the apoptosis process i.e., disappearance of mitochondria from the cell, chromatin condensation and margination, paracentral localization of nucleoli, and vacuolated nuclei. In more advanced stages of apoptosis, cells presented significant nuclear and cytoplasmic changes. It was concluded that this is the first report targeting the mitochondria, by performing inexpensive histological staining techniques, in order to assess dead cells in situ.     

Increased expression and release of functional tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) by T cells from lupus patients with active disease

Increased expression of TRAIL in membrane-bound and soluble form in patients with systemic lupus erythematosus (SLE) has been previously reported. In this study, we characterized the upregulation of T-cell-associated and soluble TRAIL (sTRAIL) in vivo and the modulation of TRAIL expression and soluble protein release in vitro following T cell activation and IFNalpha exposure. The expression of membrane-bound TRAIL as determined by flow cytometry was higher on CD4(+) and CD8(+) T cells from lupus patients compared to controls, particularly on activated CD69(+)CD8(+) T cells. Similarly, sTRAIL levels determined by ELISA were significantly elevated in serum from patients with active SLE and correlated with levels of IFNalpha. In vitro, both T-cell-associated and sTRAIL were maximally induced by T cell activation plus IFNalpha in patients and controls. By Western blot analysis, sTRAIL was detected in sera in both the monomeric and multimeric, functional form. Both forms of TRAIL were functional in vitro as determined by Annexin V staining and (51)Cr release assay but the apoptotic activity of membrane TRAIL was 2.5-fold higher compared to that of sTRAIL. These results indicate that IFNalpha-induced enhancement of TRAIL expression and of TRAIL-mediated apoptosis may amplify the abnormal apoptotic process in SLE.     

Presence of HLA-D/DR antigens on the membrane of breast tumour cells

HLA-D/DR (Ia) glycoproteins were identified in human breast carcinoma and normal mammary gland cells by means of an anti-Ia monoclonal antibody. Two techniques were used: (1) immunoperoxidase staining performed on histological sections and (2) Ia glycoproteins were isolated as follows: firstly by radioactive labelling of isolated cells, then by filtration on Sephadex G25, followed by Lens culinaris chromatography, and immune complex formation and then elution on protein A-Sepharose. Lastly, the immune complex was studied by chromatofocusing. Both techniques revealed that Ia expression was found in carcinoma cells, but not in normal cells.     

Histological evidence concerning the osseointegration of titanium implants in the fractured rabbit femur

Stabilization of the broken bone is achieved using biocompatible materials. Since histology is still considered the gold standard technique for the assessment of bone formation around metallic implants, this report investigated the titanium implant integration in the accidentally broken bone in rabbits. The experimental protocol was reviewed and approved by the Ethical Committee of the Faculty of Medicine and Pharmacy Oradea, Romania. Holes were drilled in the diaphysis of the femur, and titanium implants were inserted in the created bone defect. In two subjects, fractures occurred on days two and three after the metallic alloy implantation. The other two rabbits presented no fractures following the surgical procedure. The rabbits were euthanized and the bones (with metallic implants) were harvested for histopathological investigation. Following decalcification, the bone samples were processed using the standard paraffin technique and stained by Goldner’s trichrome procedure. In subjects with a perfect immobilization of the titanium implants, the osseointegration occurred with minimal callus formation (i.e. primary cortical healing). In rabbits with bone fractures, the callus was more exuberant. A progressive replacement of the granulation tissue with hyaline cartilage and woven bone occurred soon after. The former aspects suggested an indirect metaplasia in the created callus. In all subjects, no inflammatory cells were identified in the created callus. The bone regeneration occurred either by primary cortical healing (in perfectly immobilized titanium implants) or by a process similar to the endochondral ossification (in poorly immobilized titanium implants following accidental post-implantation bones fracture).     

Overweight and Obesity Among Hawai‘i Children Aged 4 to 5 Years Enrolled in Public Schools in 2007–2008 and Comparison with a Similar 2002–2003 Cohort

This assessment provides the most recent estimates of overweight and obesity among children 4 to 5 years old who were enrolled in public schools in the 2007–2008 school year, using data obtained from Student Health Records for 12,823 children, which represents 91% of the 14,070 children who were enrolled in kindergarten in 2007–2008. This assessment is a census of 4 to 5 year olds that entered public schools in Hawai‘i in 2007–2008 and represents approximately 38% of the total Hawai‘i population for those aged 4 to 5 years, since kindergarten is not a requirement. A limited data set with data on age, sex, height, and weight was used to calculate BMI (body mass index) percentiles. We compare this data with age and sex-specific BMI obtained from Student Health Records from 10,199 children aged 4 to 5 years entering public schools during 2002–2003. The results illustrate that like the 2002–2003 data (28.5%), over one in four (28.6%) of the children aged 4 to 5 years entering Hawai‘i public schools in 2007–2008 were either overweight or obese. Total proportions overweight and obese were persistently higher (32.5% or more in both 2002–2003 and 2007–2008) in some specific school complexes on O‘ahu as well as in some rural and Neighbor Island school complexes. Physicians, public health and school health professionals, advocates, schools, and communities should be vigilant about this persistent problem and seek to implement practices to combat overweight and obesity. In addition, the use of Student Health Records for on-going pediatric obesity surveillance should be explored more fully.     

Twelve new microsatellite loci for the shortbeard grenadier (Coryphaenoides brevibarbis)

We isolated 12 new microsatellites for the deep-sea fish Coryphaenoides brevibarbis. These loci are highly polymorphic, with allele number ranging from 2 to 17 and heterozygosity ranging from 0.231 to 0.938. These new loci will aid in identifying management units for this species.