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Cannabinoids are the active chemical components of Cannabis sativa (marijuana). The medical use of cannabis goes back over 5,000 years. Cannabinoids produce a very wide array of central and peripheral effects, some of which may have beneficial clinical applications. The discovery of cannabinoid receptors has spawned great interest within the pharmaceutical industry with the hopes of capitalizing on the beneficial effects of cannabis without the unwanted psychotropic effects on the central and peripheral nervous system. This chapter presents an overview of the pharmacology of cannabinoids and their derivatives. It reviews the current literature on central and peripheral cannabinoid receptors as related to effects on the lower urinary tract and the role of these receptors in normal and abnormal urinary tract function. An objective evaluation of the published results of clinical trials of cannabis extracts for the treatment of bladder dysfunction resulting from multiple sclerosis is also presented. It is clear that cannabinoid receptors are present in the lower urinary tract as well as spinal and higher centers involved in lower urinary tract control. Systemic cannabinoids have effects on the lower urinary tract that may be able to become clinically useful; however, a much greater understanding of the mechanisms of cannabinoid receptors in control of the human lower urinary tract is necessary to facilitate development of novel cannabinoid drugs for treatment of pelvic disorders. [ABSTRACT FROM AUTHOR]

Primary peritoneal tumors and tumor-like conditions of secondary mullerian system include various endometrioid, serous, and mucinous lesions. Endometriosis is the most common tumor-like condition resulting in a significant morbidity in women of reproductive years of age. In addition, preneoplastic hyperplasia and carcinomas (clear cell and endometrioid carcinomas) may arise from endometriosis. Primary peritoneal serous carcinoma is a frequent differential diagnosis from that of the ovarian primary. Diffuse involvement of surfaces of various abdominal organs and minimal (<5 mm in size) ovarian parenchymal involvement are diagnostic criteria for a peritoneal serous carcinoma. Various serous and mucinous cytadenomas and borderline tumors may present as extraovarian primaries. Metastatic carcinomas are the most common tumors involving the peritoneum. Gestational trophoblastic tumors primarily involving the peritoneum are extremely rare, including gestational choriocarcinoma and intermediate trophoblastic tumor (PSTT and ETT, see Chap. 8). [ABSTRACT FROM AUTHOR]

A variety of histochemical and immunohistochemical methods, if employed in the appropriate context and in a suitable combination, are useful for identifying and classifying cutaneous neural tumors. [ABSTRACT FROM AUTHOR]

We have demonstrated the synthesis of insulin by neurons within the central nervous system in vivo and in vitro. We showed that neuronal insulin induces neuronal differentiation and axonal growth. In insulin type 1 and type 2 diabetes mellitus, one major complication is nervous system neuropathy. Lack of insulin has been postulated to be the cause of the pathological alterations seen in diabetic neuropathy and diabetes mellitus has also been associated with increased risk for Alzheimer΄s disease and other types of dementia. We studied the role of insulin in the regulation of neurofilament and tau phosphorylation in vivo and in vitro. Neuronal insulin was shown to regulate neurofilament and tau phosphorylation via mitogen activated protein kinase and c-Jun N-terminal kinase. Lack of neuronal insulin promoted hyperphosphorylation of tau and neurofilament in vivo by increasing c-Jun N-terminal kinase activity and decreasing mitogen activated protein kinase. In vitro, the lack of insulin action or inhibition of mitogen activated protein kinase promoted alterations in the nonphosphorylated and phosphorylated neurofilament within the axons. Thus, insulin needs a constant presence to have a role on neurofilament and tau metabolism. Pancreatic insulin secretion is induced by high blood glucose concentration in blood, has a short half life, and the crossing of the blood brain barrier is regulated by receptors. This would make pancreatic insulin a difficult source to regulate the neuronal cytoskeleton. In conclusion, de novo synthesis of insulin by neurons is the way in which the brain has a proper continual source of insulin. These studies are also in favor of insulin΄s action in the brain and possible applications in patients with diabetes mellitus and Alzheimer΄s disease. [ABSTRACT FROM AUTHOR]

Electron microscopic immunocytochemistry takes advantage of the increased resolution of the electron microscope to localize label to specific cellular organelles. Optical microscopes limit resolution to the wavelength of light, about 0.2 μm or 200 nm. The electron microscope has theoretical resolution of less than 0.2 nm. Practically for biological samples, resolution is limited to about 2 nm because of the epoxy resin embedding. Polymers in the epoxy resin scatter the electron beam and limit the resolution of cellular structures. [ABSTRACT FROM AUTHOR]

Most of the immunocytochemistry problems seen in a shared use microscope facility are problems with the immunocytochemical procedure. In immunocytochemistry problems will frequently occur and need to be solved by troubleshooting. Most of the method problems come from three sources: (1) modified immunocytochemistry protocol that was not totally re-evaluated after being modified; (2) protocol designed without sufficient thought, generally by a novice; and (3) procedure errors where the protocol was not followed. [ABSTRACT FROM AUTHOR]

With the immunocytochemistry protocol completed, the tissue or cell preparation is now ready to be examined with a microscope. The type of microscope and the location of the microscope were determined in Chapter 9 when the Immunocytochemistry Experimental Design Chart and Section 6, Microscopy, were completed. [ABSTRACT FROM AUTHOR]

The real power of immunocytochemistry is its utility to determine whether several different proteins are colocalized under specific experimental conditions. Therefore, most immunocytochemistry experiments use multiple 1° antibodies. [ABSTRACT FROM AUTHOR]

Using 1° antibodies made in the different species is the easiest way to localize multiple proteins (Chapter 11). However, it is not possible to get all 1° antibodies needed from a different species. This is especially true because there are so many mouse monoclonal antibodies available. Eventually, an experiment will need two mouse 1° antibodies or two rabbit 1° antibodies. [ABSTRACT FROM AUTHOR]

Designing an experiment with a single 1° antibody involves many of the topics presented in the previous chapters, including reagents, types of label for antibodies, and the methods for applying antibodies. This chapter presents a step-by-step approach to designing an immunocytochemistry experiment and depends on information in the Immunocytochemistry Experimental Design Chart completed in Chapter 9. [ABSTRACT FROM AUTHOR]

In planning an immunocytochemistry experiment, decisions must be made about the exact method needed. The possibilities discussed in this chapter support a complete planning strategy for the experiment. Initially, the scientist will explore different application methods with different label types, then select for a single method and a label. The next step in the planning process is determining the details of finding and preparing reagents. The Immunocytochemistry Experimental Design Chart (included in this chapter) lists the choices that drive the method selection. Filling out the chart helps guide decisions relative to the details of the immunocytochemistry experiment and pin down the final procedure. Generating the step-by-step procedure for the experiment follows in the next chapter (Chapter 10, Single Antibody Procedure). Decisions made based on the chart in this chapter are needed before designing a first immunocytochemistry procedure. [ABSTRACT FROM AUTHOR]

Here is the fundamental issue: Can the labeling in immunocytochemistry images be trusted? Does the antibody location reveal the antigen being investigated? It is easy to think that a micrograph is naturally correct, because how would the label end up in the wrong place? Scientists experienced with immunocytochemistry accept the validity of localization, but only when appropriate controls are performed. Too often, initial excitement from an apparent positive result is dashed when control experiments show the labeling was not specific. Control experiments are essential to rule out the possibility of a nonspecific labeling being identified as specific labeling. In this chapter, three types of control protocols are presented. Together, these protocols are necessary to validate the specificity of labeling and eliminate the consideration of labeling as a misrepresentation of normal localization. [ABSTRACT FROM AUTHOR]

There many different ways to use labels or tags attached to antibodies for immunocytochemistry. Initially, in 1942, the label was attached directly to the antibody, which bound to the antigen. Since then, many different methods have been developed to indirectly bind labeled antibodies to the antigen. More recent methods have increased sensitivity in detecting low frequency antigens. [ABSTRACT FROM AUTHOR]

For years, microscopists have wanted specific stains or labels for different types of cells and different parts of cells. Prior to the mid-1900 s, only dyes were available, many of which stained special categories of compounds. Examples of dyes are hematoxylin that labels nuclei blue and eosin that labels cytoplasmic proteins pink. However, these did not allow labeling of unique proteins or compounds. The first demonstration of a method to label specific proteins came in 1942, when Albert Coons and his colleagues introduced antibody labeling of bacteria within infected human cells (Coons et al., 1942). Coons et al. used an antibody labeled with a fluorescence compound. For the first time, label conjugated to antibodies was used to specifically localize molecules in a microscope. This was the beginning of immunocytochemistry. Today, the strategies for labeling antibodies have continued to evolve and now, many labeling options are available. [ABSTRACT FROM AUTHOR]

For immunocytochemistry, fixed tissue must be cut in thin sections to be viewed in the light or fluorescence microscope. There are two common ways of sectioning tissue – the cryostat for fixed frozen tissue and the rotary microtome for paraffin-embedded tissue. In animal research, select the sectioning method based on the experimental design. [ABSTRACT FROM AUTHOR]

There are two key elements in immunocytochemistry: (1) to allow binding of the antibodies only to appropriate sites and (2) to allow antibodies access to antigens inside cells. Antibodies are proteins and can bind to tissue in ways other than specific via Fab binding. If the antibodies bind nonspecifically, then the labeling will occur as if the antibodies were bound to an antigen. Blocking of these nonspecific sites is an important and necessary step. A second requirement is that tissues and cells must be treated to allow antibodies to get inside the membranes to the cytoplasm. This is called permeabilization of cells and is done with detergents. [ABSTRACT FROM AUTHOR]

Sample preparation is one of the most important steps in immunocytochemistry because it generally receives the least amount of planning. Unfortunately, few researchers understand how critical the first few steps in an immunocytochemistry experiment are. In fact, the quality of the cells and tissue and the ability to get good results are totally dependent on initial fixation. To put sample preparation in perspective, perhaps the best thing to do is focus on the conclusion of the experiment – the quality of final microscopic image. Good images only come from cells and tissues that are fixed properly. [ABSTRACT FROM AUTHOR]

An antibody (Ab) is the key reagent of immunocytochemistry. To use antibodies effectively, consider their structure, function, and generation. Such basic knowledge about antibodies is essential to succeed in identifying suitable experimental design, finding antibodies, and trouble-shooting problems. [ABSTRACT FROM AUTHOR]

Immunocytochemistry is the use of antibodies for identifying proteins and molecules in cells and tissues viewed under a microscope. Immunocytochemistry harnesses the power of antibodies to give highly specific binding to unique sequences of amino acids in proteins. Perhaps the most exciting part of using antibodies is that new antibodies can be generated on an as-needed basis, thus providing a constant source of new reagents. Scientists are constantly generating new antibodies to specific parts of molecules thus driving continual evolution of immunocytochemistry. Identifying the location of antibodies in cells is based on availability of labels that is, itself, rapidly advancing. As time passes, immunocytochemistry continues to respond to new development of labels and advanced methods of labeling molecules. [ABSTRACT FROM AUTHOR]

In 1895, Aldred Warthin, M.D., a pathologist with a keen interest in patients, and a good listener, noted that his seamstress appeared to be depressed. He pursued this in detail, and she told him it was because she believed she would die of cancer at an early age, since everyone in her family seemed to succumb to cancer of the colon or female organs. Just as she predicted, she developed endometrial cancer and died of that disease. This brief background piqued Warthin΄s interest and he developed her pedigree and many others. In a remarkably similar scenario, Henry Lynch, M.D., in 1962, while a first year internal medicine resident, was called to see a patient who was recovering from the delirium tremens. In a statement similar to that of Warthin΄s seamstress, the patient stated that he drank because he knew he would die of cancer since everyone in the family died of colorectal cancer. Lynch, assuming that he was dealing with familial adenomatous polyposis, developed the pedigree only to find that the colon cancers were occurring in the absence of multiple colonic adenomas. Other cancers, particularly the endometrium and ovary, occurred throughout the extended family, showing a pattern consonant with an autosomal dominant mode of genetic transmission. The syndrome in the family, along with a strikingly similar family, were published in the Archives of Internal Medicine in 1966. Since that report, many hundreds of hereditary cancer-prone families with the same patterns of cancer occurrences, now known as Lynch syndrome, have been identified throughout the world, and it is now the undisputed most common hereditary form of colorectal cancer. History shows that these discoveries were products of collecting detailed family histories and innovative reasoning concerning their clinical significance. This manuscript will show the historical development of our understanding about the importance of a comprehensive cancer family history involving cancer of all anatomic sites, genetic counseling, and DNA testing when indicated. The fervent hope continues that these practices will significantly reduce cancer΄s morbidity and mortality in the Lynch syndrome. [ABSTRACT FROM AUTHOR]

Adenovirus infection is second only to rotavirus as a cause of childhood diarrhea and is associated with a broad spectrum of clinical disease in both children and adults. However, it has gained attention in recent years as a cause of diarrhea in immunocompromised patients, especially those with AIDS and patients who have undergone bone marrow or solid organ transplants. The incidence of adenovirus infection in bone marrow transplant patients has been reported at 5–20%; the incidence is lower in solid organ transplant patients. Pediatric transplant patients are affected more often than adults. Patients with severe combined immunodeficiency are also susceptible to gastrointestinal adenovirus infection. Virtually all patients present with diarrhea, sometimes accompanied by fever, weight loss, and abdominal pain. Risk factors for disseminated adenovirus infection include presence of concomitant graft-versus-host disease, use of immunosuppressive therapy, isolation of virus from multiple sites, and HLA-mismatched or unrelated transplants. [ABSTRACT FROM AUTHOR]

Herpes simplex virus (HSV) is a member of the herpesvirus group that also includes varicella zoster virus, cytomegalovirus, Epstein–Barr virus, and human herpesviruses types 6 and 8 (see also Chapters 20, 22, and 26). HSV infection may occur throughout the gastrointestinal tract, but is most common in the esophagus and anorectum. HSV infections are often seen in immunocompromised patients, but they are not limited to this group. In immunocompetent patients, infection is often self-limited; immunocompromised persons may be at risk for dissemination and life-threatening illness. Herpes simplex virus remains one of the most common infections in persons with HIV. [ABSTRACT FROM AUTHOR]

Cytomegalovirus (CMV) is best known as an opportunistic pathogen in the context of a suppressed immune system; this includes patients with AIDS and other immunodeficiency disorders, solid organ and bone marrow transplant patients, those taking chronic immunosuppressive medications, and those with underlying malignancies. CMV is the most common gastrointestinal pathogen overall in patients with AIDS, and infection can develop anywhere in the gastrointestinal tract from mouth to anus. Symptoms vary with the immune status of the patient and the site of infection; the most common clinical symptoms of gastrointestinal infection are diarrhea (either bloody or watery), abdominal pain, fever, and weight loss. Patients with esophageal infection often have dysphagia and odynophagia. [ABSTRACT FROM AUTHOR]

A high degree of clinical suspicion in the context of a great diversity of clinical symptoms remains essential to make the diagnosis of amyloidosis that still relies on tissue biopsy. Standard tissue staining should be completed with Congo red as this stain gives a specific ˵apple-green birefringence″ for amyloid under polarization. Identifying the type of amyloidosis is crucial to determine clinical management, prognosis, and treatment and thus explains that the actual nomenclature of amyloid disorders is based on the biochemical nature of amyloid. This typing requires a confrontation of clinical data to the results of some crucial blood tests such as the measure of the acute phase response and the search for a monoclonal component, histology, and genetics. Standard histological analysis is enriched with immunohistochemistry which analyses the fixation by amyloid of a panel of antibodies directed against most of the common human amyloid proteins. Rarely, sophisticated biochemical methods help to establish the true nature of the deposits. Once the steps have been cleared the disease can be classified as localized or multisystemic and its nature established. The main categories of multisystemic amylosis are the immunoglobulinic variety (AL, A for amyloid and L for light chain of immunoglobulin); amyloid associated with inflammation (AA, A for associated); and the transythyretin amyloidosis (ATTR, TTR for transthyretin) that consists in two varieties, the common ˵senile″ and the rare hereditary. [ABSTRACT FROM AUTHOR]

In situ hybridization (ISH) associated to immunohistochemistry has become a powerful tool for the examination of gene expression. Application of these techniques in grapevine tissues is limited mainly because of technical difficulties with this plant material. Here we present detailed protocols for ISH and immunohistochemistry, recommended controls and troubleshooting, along with examples of several applications to grapevine tissues. [ABSTRACT FROM AUTHOR]

Prostate-specific antigen (PSA) is the most commonly used circulating protein biomarker, which guides the diagnosis and treatment of prostate cancer. However, limitations in sensitivity and specificity limit utility of PSA when used as a diagnostic marker in screening population or as a prognostic, theragnostic, or surrogate biomarker in patients with recurrent or advanced prostate cancer. Accordingly, there is a need to discover new biomarkers to improve diagnosis, risk stratification, and therapeutic monitoring in prostate cancer. Proteomics, as an emerging technology, offers great promise in providing the cancer research community with biomarkers to guide therapeutic decision-making. However, caution must be used as new biomarkers are discovered and subject to appropriate validation before they can be applied to mainstream clinical settings. [ABSTRACT FROM AUTHOR]

Neuroendocrine tumors stand alone among other members of the larger family of tumors of the lung and upper airways. While relatively uncommon, they have, over the years, been the subject of considerable attention and study. A recent search of the literature yielded thousands of citations for small cell carcinoma, a single entity within the spectrum of neuroendocrine tumors. This chapter discusses the intriguing putative role of neuroendocrine cell hyperplasia as a possible precursor of these tumors. In addition, this chapter explores the similarities as well as differences between neuroendocrine and non-neuroendocrine tumors. Lastly, this chapter attempts to provide concise up-to-date information on both current, consensus –based morphologic criteria and the application of modern techniques, particularly immunohistochemical techniques that may facilitate histopathological diagnosis. [ABSTRACT FROM AUTHOR]

Adrenal cortical tumors may present clinically with evidence of excess hormone secretion, local mass effects, or symptoms and signs associated with malignancy. They are now also identified incidentally when the abdomen is scanned in the investigation of other disease. Although adrenal cortical carcinoma is rare, it is important to recognize it as an aggressive tumor. This chapter discusses first the aspects of the normal adrenal gland and clinical and epidemiological features of adrenal cortical tumors. The pathologic features of benign and malignant tumors are described, also the application of immunohistochemistry in differential diagnosis and an outline of the molecular pathogenesis. [ABSTRACT FROM AUTHOR]

Anterior pituitary tumors are clonal proliferation of pituitary cells. They usually consist of one cell type, although some adenomas consist of more than one cell type. Pituitary tumors can be characterized by broad spectrum markers such as synaptophysin and chromogranin. Reticulin histochemical staining is useful in separating normal hyperplastic and neoplastic pituitary tissues. Electron microscopy is a powerful tool to help separate various subtypes of adenomas including sparsely and densely granulated growth hormone adenomas and different subtypes of silent ACTH adenomas. The major types of pituitary adenomas include GH, PRL, ACTH, TSH, gonadotroph (FSH/LH), and null cell adenomas. A combination of hematoxylin and eosin staining, immunohistochemistry, and electron microscopic studies are the most comprehensive ways of classifying pituitary tumors. [ABSTRACT FROM AUTHOR]

Fine-needle aspiration biopsy (FNAB) has proved to be an excellent diagnostic tool in the initial management of various lesions affecting endocrine organs. Many studies have attested to its high sensitivity and specificity in diagnosing endocrine tumors, especially that of thyroid gland. However, as with other diagnostic tests its effectiveness is highly dependent upon the expertise of the operator performing the procedure, adequacy of the specimen, and interpretation of the cytomorphologic features. This chapter will address both old and new of the endocrine cytology: including brief description of cytologic features and differential diagnosis of various endocrine lesions and new techniques that have shown promise in their diagnosis [ABSTRACT FROM AUTHOR]

The cytopathologic characteristics of endocrine tumors occurring in non-endocrine organs are not dissimilar to their counterparts commonly found in endocrine organs and detailed in the first half of this chapter. The terminology may be organ specific such as `Merkel cell carcinoma΄ for neuroendocrine carcinoma native to the skin, but distinguishing Merkel cell from a metastatic small cell carcinoma of the lung or a small cell neuroendocrine carcinoma of paranasal sinuses may be difficult without knowledge of the clinical history and a selective immunohistochemical panel in appropriate cases. This chapter outlines the cytologic characteristics of the spectrum of neuroendocrine tumors occurring in non-neuroendocrine organs that are likely to be encountered in clinical practice by the cytopathologist. [ABSTRACT FROM AUTHOR]

Immunohistochemistry allows specific proteins to be visualized while retaining cellular or tissue structure. Sensitivity of the technique has improved over the years from a single layer of labeled antibody to amplified, labeled polymer-based systems, which enable low levels of antigen to be detected in different types of tissue preparation. However, alongside selecting the most appropriate methods of antigen recovery and enhanced signal amplification is the need to show specificity. As described in this chapter, pertinent and reliable control material must be used to both optimize and monitor performance of novel breast-associated antibodies. [ABSTRACT FROM AUTHOR]

An abstract of the article "Vomeronasal Versus Olfactory Epithelium: Is There a Cellular Basis for Human Vomeronasal Perception?," by Martin Witt and Thomas Hummel is presented.

A definition of the term "immunohistochemical staining" is presented. It is a technique which utilizes monoclonal antibodies to detect antigens on tissue sections allowing, for instance, the identification of lymphocyte subtypes.

A definition of the term "immunohistochemistry" is presented. It refers to the evaluation of protein expression/localization by incubation with antibodies that bind to specific epitopes in the protein.

A definition of the term "carcinoembryonic antigen" is presented. It refers to a cell surface glycoprotein detectable immunohistochemically in limited amounts in benign cells. It is more expressed in fetal and some neoplastic cells.