实验动物总则

We describe the development of a flow-cytometric-based opsonophagocytosis (OP) assay that is capable of looking at antigen-specific antibodies, that is, complement-dependent, not complement-mediated. Using the OP assay we are able to look at uptake of bacteria by neutrophils simul ...

The interaction of bacteria with mucosal surfaces of the respiratory tract is thought to be critical in their pathogenesis. Bacteria must overcome local defenses that include physical barriers such as mucus, beating cilia, and an intact epithelial surface; antibacterial substances ...

Haemophilus influenzae type b, a colonizer of the human respiratory tract, is capable of breaching the epithelial cellular barrier of this niche, as well as endothelial barriers of the vasculature to enter the blood system, prior to dissemination to other tissues including the brain (1). Besi ...

Isolates of Haemophilus influenzae can be separated into encapsulated and nonencapsulated forms. Encapsulated strains express one of six structurally and antigenically distinct capsular polysaccharides, designated serotypes a–f (1). In contrast, nonencapsulated str ...

Haemophilus influenzae is a Gram-negative bacterium that was first described by Pfeiffer in 1892 (1). This ubiquitous, human-specific organism was originally thought to be the etiologic agent of “influenza.” However, H. influenzae was not consistently isolated from autopsied lungs ...

Although Haemophilus influenzae genes may be transferred by plasmidmediated conjugation, this process is not discussed further below, as it has received little attention and is rarely used for strain construction. Conjugative plasmids are common in H. influenzae, as in other bacter ...

Haemophilus influenzae is differentiated from other Haemophilus species primarily by its growth requirements for both hemin (called factor X in the old literature) and nicotinamide adenine dinucleotide (NAD or factor V). H. influenzae strains normally grow well in rich media such as br ...

With the advent of effective vaccination strategies, life-threatening Haemophilus influenzae infections are now rare in developed countries (1). However, H. influenzae can still cause infection at a variety of anatomical sites, especially the upper and the lower airway. For this reas ...

The bacterial proteome is the total protein complement expressed from the genome under defined conditions of growth. Unlike the genome, the proteome is not a fixed characteristic, but varies with the conditions of growth of the organisms from which proteins are harvested. Proteomics gives ...

Antibiotic resistant strains of Haemophilus influenzae appeared in the early 1970s and studies on these strains led to the detection of large 40–50 kb conjugative plasmids (1–3). Early work was hampered by the failure to easily detect extrachromosomal plasmid in clinical isolates. Howe ...

A single phenotypic change observed during an organism’s growth or adaptation to its environment is usually the result of a coordinated expression of genes ranging from a few to many. Various techniques are in use today for quantifying gene expression. These include indirect methods, such as p ...

Reporter genes are widely used in molecular biology. In mammalian cells, expression of a gene encoding luciferase allows the monitoring of promoter activity through the measurement of light produced during catalysis. In yeast and procaryotic cells, the lacZ gene of Escherichia coli (en ...

This chapter outlines the generation and application of human papillomavirus type 33 (HPV 33) pseudovirions. These pseudovirions are structurally indistinguishable from native virions and are therefore valuable tools for the study of papillomavirus/cell interactions. The ...

Hematoxylin and eosin (H&E) staining of cutaneous warts is presented to illustrate the practical methods utilized for histochemical analysis of cutaneous human papillomavirus-associated lesions. Every step of the staining procedure, from sampling of the specimens to microsc ...

A wide interobserver variation is seen even among competent histopathologists in the routine diagnosis of cervical intraepithelial neoplasia (CIN). As a result, early detection of low-grade CIN (CIN 1) lesions, in particular, remains a major challenge both in routine diagnosis and in ce ...

Human papillomaviruses (HPVs) are known to be etiological agents of cervical cancer and have been found in 99.7% of women with high-grade (HG) cervical intraepithelial neoplasia (CIN) precancer. Testing of high-risk HPV (HR-HPV) has been proposed as a way of improving cervical screening, es ...

The key events during the papillomavirus life cycle can be mapped in infected tissue samples by antibody detection and in situ hybridization. The ease of immuno-detection varies for different proteins and is dependent on antigen availability. Epitope exposure is sometimes necessary, ...

Most cervical cancers are preventable when the precursor lesions are detected in time. Human papilloma viruses (HPVs) are the main risk factors for cervical cancer development, but there is a high percentage of healthy women infected with HPV that never develop a lesion. Only a small percentage ...

The treatment of cervical intraepithelial neoplasia (CIN) depends on the evaluation of CIN grade. The grading of CIN is however problematic, as intra- and interobserver reproducibility of CIN-grade evaluation among pathologists is not perfect. There are also difficulties in reliab ...

Polymerase chain reaction (PCR)-based assays that use consensus primers to detect DNA of a broad spectrum of human papillomavirus (HPV) types in a single assay belong to the most frequently used methods to detect HPV in clinical specimens. Here, we describe in detail one of these assays, the so-cal ...