From: owner-ammf-digest@smoe.org (alt.music.moxy-fruvous digest) To: ammf-digest@smoe.org Subject: alt.music.moxy-fruvous digest V14 #4456 Reply-To: ammf@fruvous.com Sender: owner-ammf-digest@smoe.org Errors-To: owner-ammf-digest@smoe.org Precedence: bulk alt.music.moxy-fruvous digest Saturday, June 27 2020 Volume 14 : Number 4456 Today's Subjects: ----------------- It involves a clever Netflix loophole that may change your life ["Perpetu] SHOOTERS: This thing aims the gun for you (Hurry!) ["Shooting Gear" Subject: It involves a clever Netflix loophole that may change your life It involves a clever Netflix loophole that may change your life http://completes.today/wTsVn1DOnqcowQPUFpwGwTEpOiMSIDpRnVWRXVjGoBEfXVg http://completes.today/EMsfy1jakOEfH1qFu54Tq2TDJng2bo71dKX6NO0Xl0eE3X2M To perform in vitro analysis, a protein must be purified away from other cellular components. This process usually begins with cell lysis, in which a cell's membrane is disrupted and its internal contents released into a solution known as a crude lysate. The resulting mixture can be purified using ultracentrifugation, which fractionates the various cellular components into fractions containing soluble proteins; membrane lipids and proteins; cellular organelles, and nucleic acids. Precipitation by a method known as salting out can concentrate the proteins from this lysate. Various types of chromatography are then used to isolate the protein or proteins of interest based on properties such as molecular weight, net charge and binding affinity. The level of purification can be monitored using various types of gel electrophoresis if the desired protein's molecular weight and isoelectric point are known, by spectroscopy if the protein has distinguishable spectroscopic features, or by enzyme assays if the protein has enzymatic activity. Additionally, proteins can be isolated according to their charge using electrofocusing. For natural proteins, a series of purification steps may be necessary to obtain protein sufficiently pure for laboratory applications. To simplify this process, genetic engineering is often used to add chemical features to proteins that make them easier to purify without affecting their structure or activity. Here, a "tag" consisting of a specific amino acid sequence, often a series of histidine residues (a "His-tag"), is attached to one terminus of the protein. As a result, when the lysate is passed over a chromatography column containing nickel, the histidine residues ligate the nickel and attach to the column while the untagged components of the lysate pass unimpeded. A number of different tags have been developed to help researchers purify specific proteins from complex mixtures. Cellular localization Proteins in different cellular compartments and structures tagged with green fluorescent protein (here, white) The study of proteins in vivo is often concerned with the synthesis and localization of the protein within the cell. Although many intracellular proteins are synthesized in the cytoplasm and membrane-bound or secreted proteins in the endoplasmic reticulum, the specifics of how proteins are targeted to specific organelles or cellular structures is often unclear. A useful technique for assessing cellular localization uses genetic engineering to express in a cell a fusion protein or chimera consisting of the natural protein of interest linked to a "reporter" such as green fluorescent protein (GFP). The fused protein's position within the cell can be cleanly and efficiently visualized using microscopy, as shown in the figure opposite. Other methods for elucidating the cellular location of proteins requires the use of known compartmental markers for regions such as the ER, the Golgi, lysosomes or vacuoles, mitochondria, chloroplasts, plasma membrane, etc. With the use of fluorescently tagged versions of these markers or of antibodies to known markers, it becomes much simpler to identify the localization of a protein of interest. For example, indirect immunofluorescence will allow for fluorescence colocalization and demonstration of location. Fluorescent dyes are used to label cellular compartments for a similar purpose. Other possibilities exist, as well. For example, immunohistochemistry usually utilizes an antibody to one or more proteins of interest that are conjugated to enzymes yielding either luminescent or chromogenic signals that can be compared between samples, allowing for localization information. Another applicable technique is cofractionation in sucrose (or other material) gradients using isopycnic centrifugation. While this technique does not prove colocalization of a compartment of known density and the protein of interest, it does increase the likelihood, and is more amenable to large-scale studies. Finally, the gold-standard method of cellular localization is immunoelectron microscopy. This technique also uses an antibody to the protein of interest, along with classical electron microscopy techniques. The sample is prepared for normal electron microscopic examination, and then treated with an antibody to the protein of interest that is conjugated to an extremely electro-dense material, usually gold. This allows for the localization of both ultrastructural details as well as the protein of interest. Through another genetic engineering application known as site-directed mutagenesis, researchers can alter the protein sequence and hence its structure, cellular localization, and susceptibility to regulation. This technique even allows the incorporation of unnatural amino acids into proteins, using modified tRNAs, and may allow the rational design of new proteins with novel properties ------------------------------ Date: Thu, 25 Jun 2020 05:55:22 -0400 From: "Shooting Gear" Subject: SHOOTERS: This thing aims the gun for you (Hurry!) SHOOTERS: This thing aims the gun for you (Hurry!) http://alivehear.us/RCZ7xGThrPXZHNC5jtb6dCdFNarAQLNSE_alBTmv6MqmOqke http://alivehear.us/K0Ov6PooULKgJdf462Mgby7rmaSF49v8i9YtByfRCOOVBHTD formal rules to define right and wrong. Girls, on the other hand, have a care and responsibility perspective where personal relationships are considered when judging a situation. Gilligan also studied the effect of gender on self-esteem. She claimed that society's socialization of females is the reason why girls' self-esteem diminishes as they grow older. Girls struggle to regain their personal strength when moving through adolescence as they have fewer female teachers and most authority figures are men. As parents are present in a child's life from the beginning, their influence in a child's early socialization is very important, especially in regards to gender roles. Sociologists have identified four ways in which parents socialize gender roles in their children: Shaping gender related attributes through toys and activities, differing their interaction with children based on the sex of the child, serving as primary gender models, and communicating gender ideals and expectations. Sociologist of gender R.W. Connell contends that socialization theory is "inadequate" for explaining gender, because it presumes a largely consensual process except for a few "deviants," when really most children revolt against pressures to be conventionally gendered; because it cannot explain contradictory "scripts" that come from different socialization agents in the same society, and because it does not account for conflict between the different levels of an individual's gender (and general) identity. Racial socialization Racial socialization, or racial-ethnic socialization (Q96195578), has been defined as "the developmental processes by which children acquire the behaviors, perceptions, values, and attitudes of an ethnic group, and come to see themselves and others as members of the group". The existing literature conceptualizes racial socialization as having multiple dimensions. Researchers have identified five dimensions that commonly appear in the racial socialization literature: cultural socialization, preparation for bias, promotion of mistrust, egalitarianism, and other. Cultural socialization refers to parenting practices that teach children about their racial history or heritage and is sometimes referred to as pride development. Preparation for bias refers to parenting practices focused on preparing children to be aware of, and cope with, discrimination. Promotion of mistrust refers to the parenting practices of socializing children to be wary of people from other races. Egalitarianism refers to socializing children with the belief that all people are equal and should be treated with a common humanity. Oppression socialization Oppression socialization refers to the process by which "individuals develop understandings of power and political structure, particularly as these inform perceptions of identity, power, and opportunity relative to gender, racialized group membership, and sexuality." This action is a form of political socialization in its relation to power and the persistent compliance of the disadvantaged with their oppression using limited "overt coercion." Language socialization Based on comparative research in different societies, focusing on the role of language in child development, linguistic anthropologists Elinor Ochs and Bambi Schieffelin have developed the theory of language socialization. They discovered that the processes of enculturation and socialization do not occur apart from the process of language acquisition, but that children acquire language and culture together in what amounts to an integrated process. Members of all societies socialize children both to and through the use of language; acquiring competence in a language, the novice is by the same token socialized into the categories and norms of the culture, while the culture, in turn, provides the norms of the use of language. ------------------------------ End of alt.music.moxy-fruvous digest V14 #4456 **********************************************