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Apoptosis: Molecular Regulation of Cell Death and Hematologic Malignancies

Apoptosis, or programmed cell death, represents in cell biology a functional program as important as cell growth or differentiation. Programmed cell death is of basic importance for the development of multicellular organisms and its basic mechanisms are conserved during the evolut ...

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Cytokine Receptors: Qualitative and Quantitative Analysis

Much has been written on the possible role of cytokines in hematopoietic malignancies. In some instances, such as interleukin-6 (IL-6) in multiple myeloma, there is evidence for a central role in maintaining malignant growth (1). The relationship between IL-6 and multiple myeloma is probab ...

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A Rat Pain Model of Facial Cancer

Cancer pain is a very severe problem for patients with advanced or terminal cancer. However, the induction mechanism remains unknown. Orofacial cancer patients often report difficulties in eating and swallowing, different from patients with cancer in other regions. Although sever ...

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Unilateral T13 and L1 Dorsal Root Avulsion: Methods for a Novel Model of Central Neuropathic Pain

Central neuropathic pain is associated with many disease states including multiple sclerosis, stroke, and spinal cord injury, and is poorly managed. One type of central neuropathic pain that is particularly debilitating and challenging to treat is pain that occurs below the level of inju ...

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Orofacial Pain Models and Behavior Assessment

Orofacial pain remains an understudied area in pain research given that most attention has been focused on the spinal system. In this chapter, animal models of neuropathic and inflammatory orofacial pain are presented. Four different types of pain behavior tests are then described for ass ...

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A Rat Chronic Pain Model of Spinal Cord Contusion Injury

Spinal cord injury-induced pain is a common clinical problem affecting adversely the quality of daily lives of spinal cord injured patients. Management with current pain medications can only lead to partial pain relief in some spinal cord injured patients, which is usually associated wi ...

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A Lumbosacral Ventral Root Avulsion Injury and Repair Model for Studies of Neuropathic Pain in Rats

Neuropathic pain may develop after a variety of injuries to peripheral nerves and roots. Most injury models have included a direct injury to primary afferent fibers or neurons. Recently, it has been demonstrated that injury to motor fibers in ventral roots may also result in neuropathic pain. A lu ...

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Modeling Calcium-Overload Mediated Necrosis in Drosophila

Necrotic cell death (necrosis) occurs in many acute-onset diseases. However, our poor understanding of its mechanism has greatly limited medical interventions. Here we describe two methods to establish necrosis models in Drosophila. Our strategy is to overload calcium by expression ...

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Intracytoplasmic Morphology-Selected Sperm Injection

Approximately 40% of sterility in couples can be attributed to male subfertility and intracytoplasmic sperm injection (ICSI) has become a powerful tool in assisted reproduction to overcome male infertility. Intracytoplasmic morphologically selected sperm injection (IMSI) ...

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Sperm Cryopreservation Methods

There are multiple clinical situations where cryopreservation of sperm is required including sperm banking prior to chemotherapy or radiation treatment for cancer, donor sperm for couples without a male partner, and various etiologies of male factor infertility. Bunge and Sherman ...

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Human Y Chromosome Microdeletion Analysis by PCR Multiplex Protocols Identifying only Clinically Relevant AZF Microdeletions

PCR multiplex assays are the method of choice for quickly revealing genomic microdeletions in the large repetitive genomic sequence blocks on the long arm of the human Y chromosome. They harbor the Azoospermia Factor (AZF) genes, which cause male infertility when functionally disrupted. ...

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Flow Cytometric Methods for Sperm Assessment

Flow cytometry allows the assessment of multiple sperm parameters following a diverse number of protocols. Here, we describe three methods for evaluating critical aspects of sperm quality: the double stain SYBR-14 and propidium iodide (PI) for assessing sperm viability; the double sta ...

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Sperm Aneuploidy Testing Using Fluorescence In Situ Hybridization

Sperm aneuploidy screening has been used as a tool in diagnosis and determining treatment options for male factor infertility since the development of human sperm karyotyping by injection into hamster and mouse oocytes in the 1970s. From these studies and subsequent work with interphase ...

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Methods for the Assessment of Sperm Capacitation and Acrosome Reaction Excluding the Sperm Penetration Assay

Assessing the ability of human spermatozoa to acquire fertilizing potential (capacitation) by stimulating exocytosis of the contents of the acrosome (acrosome reaction) is thought to have diagnostic potential (De Jonge, Reprod Med Rev 3:159–178, 1994). Calcium-mobilizing agen ...

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Phenotypic Assessment of Male Fertility Status in Transgenic Animal Models

This chapter describes the approach to define the cause of male infertility in a genetically modified male mouse. It provides a guide to the establishment of the infertility status and whether it is due to the failure of mating or due to abnormalities of the sperm output, motility, and morphology. Fur ...

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Methods for Sperm-Mediated Gene Transfer

The transgenic technologies represent potent biotechnological tools that allow the generation of genetically modified animals useful for basic research and for biomedical, veterinary, and agricultural applications. Among transgenic techniques, we describe here the spe ...

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Transgenic Modification of Spermatogonial Stem Cells Using Lentiviral Vectors

The continuous production of spermatazoa throughout the reproductive lifetime of a male depends on the maintenance of a pool of progenitor cells called spermatogonial stem cells (SSCs). SSCs represent a very small fraction of the cellular population in the testes and lack definitive mol ...

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Testicular Tissue Grafting and Male Germ Cell Transplantation

Testicular tissue grafting and male germ cell transplantation are techniques that offer unprecedented opportunities to study testicular function and development. While testicular tissue grafting allows recapitulation of testis development and spermatogenesis from ...

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Methods for Direct and Indirect Antisperm Antibody Testing

Antisperm antibodies (ASA) are one well-known cause of refractory infertility in both males and females. In females, a sperm immobilization test, which detects sperm-immobilizing antibodies indirectly in the patient’s serum, requires complement for the reaction and thus seems to be a m ...

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Sperm Morphology Classification: A Rational Method for Schemes Adopted by the World Health Organization

Sperm morphology is an important measure of testicular health, spermiation, and fertility potential. The World Health Organization (WHO) Semen Manuals advocate different sperm morphology schemes, but, like the schemes themselves, do not describe classification sequence or ru ...

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