Type I cytokine receptors are transmembrane receptors expressed on the surface of cells that recognize and respond to cytokines with four alpha-helical strands. These receptors are also known as hemopoietin receptors and share a common amino acid motif (WSXWS) int he extracellular portion. Type I cytokine receptors include interleukin receptors, colony stimulating factor receptors and other cytokine receptors.

The signal transducing chains are often shared between different receptors within the family. For example, the IL-2 receptor shares a common gamma chain (CD132) with the IL-4 reeptor. The common beta chain (CD131) is shared between the GM-CSF recetpor and the IL-3 receptor.

Leptin or obesity receptor (Ob-R) is a ember of class I cytokine receptor family, Ob-R, expressed in six isoforms, is the product of alternative RNA splicing of db gene. According to its structural differences, the receptor’s isoforms are divided into three classes: long, short, and secretory isoforms. A long, fully active isoform of Ob-Rb is expressed mainly in the hypothalamus, where it takes part in energy homeostasis and in the regulation of secretory organs’ activity. Ob-Rb is also present on all types of immune cells, invovled in innate and adaptive immunity. Short leptin isoforms that contain box 1 motif are able to bind JAK kinases as well as to activate some other signal transduction cascades. A solube isoform can regulate serum leptin concentraiton and serve as a carreir protein delivering the hormone to its membrane receptors and is able to transduct the signal into the cell. (Wasik, “Leptin Recetpors” European Journal of Medical Research” 2010, 50-54).

A long, fully active isoform of Ob-Rb is epxressed mainly in the hypothalamus, where it takes part in energy, homeostasis, and int he regulation of secretory organs’ activity. Ob-Rb is also rpesent in all types of immune cells, involved in innate and adaptive immunity. (Wasik, “Leptin Recetpors” European Journal of Medical Research” 2010, 50-54).

Lack of a full-lenght Ob-Rb recetpor is responsible for the development of the early obesity phenotype in db/db mice and in obese rats. (Wasik, “Leptin Recetpors” European Journal of Medical Research” 2010, 50-54).

Short leptin isoforms that comtain box 1 motif are able to bind JAK kisnases and to activate some signal transduction cascades. However, the effect of short isoform activation differs from that of long isoform activation. Their main function is presumably connected with leptin internalization and degradation. A short isoform, Ob-Ra is the most common Ob-R isoform that can be found in many various cells and tissues, cinlduign kidney, lugns, liver, speen, and macroppages. (Wasik, “Leptin Receptors” European Journal of Medical Research” 2010, 50-54).

A major role in leptin signal transduction through membrane receptors is mediated through JAK/STATE pathway. Among all Ob-R isoforms, only the full lenght isoform is able to fully transduce and activaiton signal into the cell. Ob-Rb is considered a fully active receptor, because it contains 3 intracellular motifs necessary to activate the JAK/STATE pathway.

Websites:  Grow Weed Easy

Books/References on Growing Cannabis:

Details of existing Cannabis plants varieties and breeding methods are described in Potter et al. (2011, World Wide Weed: Global Trends in Cannabis Cultivation and Its Control), Holland (2010, The Pot Book: A Complete Guide to Cannabis, Inner Traditions/Bear & Co, ISBN1594778981, 9781594778988), Green I (2009, The Cannabis Grow Bible: The Definitive Guide to Growing Marijuana for Recreational and Medical Use, Green Candy Press, 2009, ISBN 1931160589, 9781931160582), Green II (2005, The Cannabis Breeder’s Bible: The Definitive Guide to Marijuana Genetics, Cannabis Botany and Creating Strains for the Seed Market, Green Candy Press, 1931160279, 9781931160278), Starks (1990, Marijuana Chemistry: Genetics, Processing & Potency, ISBN 0914171399, 9780914171393), Clarke (1981, Marijuana Botany, an Advanced Study: The Propagation and Breeding of Distinctive Cannabis, Ronin Publishing, ISBN 091417178X, 9780914171782), Short (2004, Cultivating Exceptional Cannabis: An Expert Breeder Shares His Secrets, ISBN 1936807122, 9781936807123), Cervantes (2004, Marijuana Horticulture: The Indoor/Outdoor Medical Grower’s Bible, Van Patten Publishing, ISBN 187882323X, 9781878823236), Franck et al. (1990, Marijuana Grower’s Guide, Red Eye Press, ISBN 0929349016, 9780929349015), Grotenhermen and Russo (2002, Cannabis and Cannabinoids: Pharmacology, Toxicology, and Therapeutic Potential, Psychology Press, ISBN 0789015080, 9780789015082), Rosenthal (2007, The Big Book of Buds: More Marijuana Varieties from the World’s Great Seed Breeders, ISBN 1936807068, 9781936807062), Clarke, R C ( Cannabis: Evolution and Ethnobotany 2013 (In press)), King, J (Cannabible Vols 1-3, 2001-2006), and four volumes of Rosenthal’s Big Book of Buds series (2001, 2004, 2007, and 2011), 

Introduction/Definitions:

Typically, sun-grown Cannabis is planted in spring, flowers when night periods exceed about 10-12 hours, and is ready to harvest in late autum. 

Cannabis is a very rapidly growing plant, attaining a usual height of three to twenty feet at maturity. Cannabis is dioecious, which means that there are sexually distinct male and female plants. The known species are Cannabis sai/Va, Cannabis indica and Cannabis ruderatis with multiple strains in cultivation. 

Photoperiod refers to a plant’s response to the amount of light and darkness, to which it is exposed. Short-day or long-night plants, as obligate photoperiodic plants, will only begin flowering once the sunlight hours are reduced to a certain number, based on the seasonal changes of the earth’s orbit or artificial replication thereof. Typically, short-day plants will flower when the day is less than 12 hours (i.e., the night is longer than 12 hours) regardless of plant age or size. In indoor growing operations, this photosensitivity allows for a precisely tailored plant cycle for continuous growing seasons with the stages of development being artificially controlled. Additionally, when outdoors, short-day plants can be fooled into flowering early (i.e., outside of the natural seasonal schedule) by being covered for at least 12 hours in a 24-hour period. Similarly, if exposed to more than 12 hours of light in a 24-hour period, short-day plants will not flower, so flowering may be delayed and/or a plant may be kept in a perpetual vegetative state (e.g., as a mother plant for clones and/or seeds). (US 20230242932)

In general, a Cannabis plant has a vegetative stage and a flower stage. The latter may be initiated by substantially increasing the time the Cannabis plant essentially does not receive light (dark period). During the phase of life known as the vegetative stage (the first stage of life for marijuana), a cannabis plant grows like a weed. In the vegetative stage a cannabis plant essentially only grows new stems and leaves, and can grow several inches a day. When growing Cannabis indoors, the flowering stage begins when the lighting schedule is reduced to a 12/12 light cycle (12 hours light, 12 hours darkness each day). During the vegetative stage, the Cannabis plant should get horticulture light during at least about 16 hours a day. Getting those 12 hours of uninterrupted darkness each day may give the plant the signal that it is time to start flowering. The plant may receive a kind of winter trigger, because the days are getting short. On the internet, it is amongst others found that “if the plant gets any light during the dark period, even for just a minute, it will not make buds! A flowering plant may even revert back or express hermaphroditism if it gets any light at night”. During the first few weeks after being switched to a 12/12 schedule, the Cannabis plant will be growing relatively fast and may rapidly gain height. In fact, a Cannabis plant can almost double in height after the switch to 12/12. This period of super-fast and often stretchy growth is sometimes referred to as the “flowering stretch”. The female plants will start sprouting lots of white pistils, though they usually will not start growing “real” buds with substance quite yet. (US 20220295712)

With regard to some plants, as an example, such as cannabis, there are two key growth stages, a vegetative stage and flowing stage. To maintain the plant in the vegetative cycle, the lights may be kept ‘on’ eighteen (18) hours a day and turned ‘off’ six (6) hours a day. When the grower wants to flip the plant into the flowing stage or flower producing stage, light times may be changed to twelve (12) hours ‘on’ and twelve (12) hours ‘off’ in a 24-hour period. The change in light times activates the plant into what it perceives as a season change, thereby moving the plant into the flowing stage. US 10842082

Propagation:

In vitro Propagation:

In vitro propagation techniques offer efficient multiplication yields of disease free C. sativa L. plants at a commercial scale. Generally, such techniques eliminate cultivation space and reduce production costs and time. The products of these processes display genetic and phenotypic univformity. in terms of theri morphological trails, which include some of the majjor commerically important trails. Moreover, the method has tremendous potential for genetic transformation by modifying both the genetic information and the regulation of tose gene responsible for the production of valuable biolgoical active substances. (Mitsis, “An Alternative in vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) presenting efficient rooting, for commerical production” Plants, 2022, 11(10): 1333).

Traditionally, the conventional in vitro micropropagation prcoedure contains four stages, i.e., culture establishment, proliferation, rooting of shoots and acclimatization. In vitro rootingreported costs represent 35-75% of the total process. Gradually labor costs relating to conventional micropropagation, with the exception of culture extablishment, have reached about 60%. Cuttings, in teh frist three pahses,a re encldsed in vessels wtih apssive gas exchange, udner aseptic conditions, An upturn in tissue culture led to the onset of the phtooautotrophic micropropagation technqiue, in which chlorophyllous explants were grown under CO2-rich condtions. Moreover, culturing explants in vessels with gas-permeable film as enclosures, combined with the use of rockwool multi-blocks as a substrate were found to be suitable for the development of some plant species. Phtoautotrophic micropropagation on rockwool blocks as a substrate was efficient for C. sativa L. cultivation as well. An alternative that has further improved on teh perfromance of the vitro process is the use of a double-phase culture system (semi-solid medium with a layer of liquid medium on the top), in which shooting and rooting are performed simultaneously. Other approaches include the sue of bioreactors, which improve the physiological state of the explants. Such systems are commercailly available, such as automated tmporary immersion (RITA® and Plantform ™ bioractors, the rocker system as well as others. (Mitsis, “An Alternative in vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) presenting efficient rooting, for commerical production” Plants, 2022, 11(10): 1333).

(Mitsis, “An Alternative in vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) presenting efficient rooting, for commerical production” Plants, 2022, 11(10): 1333) discloses an in vitro propagation protocol in vitro cutting disinfecstation, culture establishment, and root induction, as well as acclimimatization of the in vitro-propagated plantlets using pet-based sponges as a substrate, impregnated in liquid medium. Root initiation of in vitro-propagated cuttings was commenced during the third week fo culture, depending on the rooting treatment. Cuttings were excised from selected healthy young medical C. sativa L. plants at the vegetative grwoth stage. Two varieties, a high-cannabidiol plant variety (H_CBD) and a high-cannabigerol plant variety (H_CBG), of C. sativa L. (Cannabaceae) were included in the study. The cuttings’ donor plants were grown in a green house. Elite (based on chemical profile) female plants were used in the xperiments. Motehr plants were selected ruing a previous research study and maintained at the vegetative stage for a photoperiod of 18 h. All plants were kept indoors, under controlled envornmental conditions at 27C ± 2 C. 

Supply/Type of Light Used:

To determine the appropriate lighting (and the best lamp to use), the specific needs of the plant must be considered, as well as the room size and ventilation. To arrange optimal lighting, the lighting present in the plant’s natural environment needs to be imitated. For example vegetables grow best in full sunlight, which means in practice that as much light as possible must be supplied to grow cannabis indoors (high intensity discharge (HID) lights such as high pressure sodium (HPS) and metal halide (MH) are preferred. Fluorescent lamps can also be used). Incandescence and mercury vapor lighting are not used in cannabis cultivation. (US 20120311744)

In addition, plants also require both dark and light (“photo”-) periods. As such, lights need to be timed to switch them on and off at set intervals. The optimum photo/dark-periods is specific depending on each plant (some prefer long days and short nights and others preferring the opposite, or something in between). Most plants will grow under most light spectra, yet always prefer a full spectrum light (HPS). However, certain plants (as cannabis) can be grown successfully under both types of light. MH is used for vegetative phase of growth, as it encourages short inter nodes (distance between sets of leaves), and inhibits cell elongation, creating a shorter, stockier plant. Metal halide lamps produce more ultraviolet radiation than high pressure sodium lamps, which may play a role in increasing the flowering (and for certain plants as cannabis the amount of working substances as THC) produced by the plant. High pressure sodium lamps trigger a greater flowering response in the plant and are thus used for the second (or reproductive) phase of the growth. If high pressure sodium lamps are used for the vegetative phase, plants will usually grow slightly more quickly, but will also have longer inter nodes, and may be taller. (US 20120311744)

According to the inverse square law, the intensity of light radiating from a point source (in this case a bulb) is inversely proportional to the square of the distance from the source. So if an object is twice as far away, it receives only 1/4 the light. This is a serious hurdle for indoor marijuana growers, and many techniques are employed to use light as efficiently as possible. (US 20120311744)

Reflectors are often used in the lamps to maximize light efficiency. Plants or lights are moved as close together as possible so that they receive equal lighting and that all light coming from the lamps wind up on the plants (rather than partly besides it). Often, the distance between lamp and plant is in the range of 0.6 m (2 ft) with incandescent lamps, to 10 cm (4 in) with other lamps, such as compact, large and high-output fluorescent lamps. Some marijuana cultivators cover the walls of their grow-room with some type of reflective material (often Mylar), or alternatively, white paint to maximize efficiency. (US 20120311744)

One commonly used covering is 6 millimeter (150 .mu.m) PVC plastic sheeting that is white on one side and black on the other. The plastic is installed with the white side facing in to the room to reflect light, and the black facing the wall, to reduce fungus and mold growth. Another common covering is flat white paint, with a high titanium dioxide content to maximize reflectivity. Mylar sheeting from a grow store is very effective when it lines grow room walls, along with Astrofoil (which also reflects heat), and Foylon (a foil-laminated, reinforced fabric). (US 20120311744)

LED: Recent advancements in LED technology have allowed for diodes that emit enough energy for cannabis cultivation. One major short coming of LED’s in the past has been a lack of intensity. Higher wattage chips are required to produce enough luminous efficiency to produce larger, denser yields. As with using a 400 w HPS vs. a 1000 w HPS, intensity has everything to do with yield. The same applies to LEDs however, it is not as simple as measuring watts because better quality chips can produce more light with less watts than cheap chips running at lower watts.LED grow lights are still considered an experimental technology in cannabis cultivation. The market remains flooded with cheap quality LED lights that do not produce yields comparable to what growers are accustomed to. Many companies are using single watt LED chips, which have notoriously produced low yields and wispy results. Growers should look for lights with 6 watt chips. When considering purchasing LED grow lights, one should carefully examine both the spectrum and the intensity of the light. The advantages of LEDs, low heat output, long life span, and simpler environmental control, coupled with the ever increasing quality of the technology ensure that they can potentially mark a significant transformation in the cultivation of cannabis. NASA has experimented with LED panel light sources on plant growth.(US 20120311744)

HPS bulb: has most of the light spectrum in the “orange” range, with almost no `blue` and very little `red.` For this reason, it is poor in the 430-460 nm, and poor in the 680-700 nm. Luckily, the light is so powerful that the spill-over at these frequencies is still sufficient to do a good job. The principal shortcoming of the HPS lamp turned it into an advantage for LEDs. LED lights allow one to focus intensity in the high PAR absorption range of the light spectrum. New models of LED grow lights incorporate multiple types of chips that cover the whole range of red light, blue light, and now full spectrum light. (US 20120311744)

The cannabis plant belongs to the genus Cannabis. There are varieties of different cannabis plant strains, which are suitable for the production of cannabis for medicinal applications. Cannabis material for medical use may be produced from dried cannabis flowers or from extracts of cannabis flowers. A cannabis flower can comprise several hundred different active ingredients, among them plenty of different terpenes and cannabinoids. One of the most active cannabinoids is tetrahydrocannabinol (THC). Other cannabinoids are for example cannabidiol (CBD), cannabinol (CBN), cannabicyclol (CBL), cannabichromene (CBC), cannabigerol (CBG) and delta-8-tetrahydrocannabinol. Terpenes produced by the cannabis plant can influence the physiological effect of cannabinoids. Cannabis material is useful in the treatment of several diseases such as multiple sclerosis. Cannabis material may also have beneficial effects in patients who suffer from chronic pain, muscle spasms, insomnia, posttraumatic stress disorder, dizziness during chemotherapy and low appetite. Cannabis material for medical use can be in the form of capsules, tinctures or edibles. (EP3771330).

The Cannabis plant produces numerous secondary metabolites including cannabinoids and terpenes, which are known for their therapeutic effect. Many of these metabolites are produced in a special structure, termed glandular trichome, developed mostly on leaves called bract that encapsulates the female’s reproductive parts in the female flower. The cola, which is a cluster of flower buds (inflorescences) that grow tightly together, develops upon induction of flowering on every growing tip, emerging from leaf nodes along the stem. As the flower develops the trichrome go through ripening process having three visible stages: translucent, opaque, and amber, simultaneously with a change in the content of the secondary metabolites.

Cannabis plants contain cannabinoids, such as tetrahydrocannabino (THC) and cannabidol (CBD), which can be eaten, inhaled, or otherwise absorbed into a person’s body for medical, spiritual, or recreational purposes. At maturity, a femal cannabis plant will include infructescences (also referred to as buds) that can have up to ten times higher levels of calnabinoids than its leaves and up to one hundred times higher level of cannabinoids that its stalks. 

What Makes a Potent Cannabis Plant?

There is much more to cannabis than THC. When it comes to moving a product, only two numbers seem to matter; the list price and its THC content. Super potent cannabis flower, with THC of greater than 25%, will dominate dispensary shelves and justify charging $75 or more. However, a study at the Unviersity of Colorado publisehd in JAMA Psychiatry found that THC content is a poor indicator of potency. High-THC weed does not ven mean one gets more high. The reason for this is that there are many more factors at play than THC. There are a host of cannabinoids, including CBD as well as more than 100 others, most of which are not even tested for. There are also aromatic compounds called terpenes that dictate how cannabis affects the mind and body. All of these work in concert, a phenomenon known as the “the entrouage effect”. This is why synthetic THC simply never had the same medical effects a smoking week. A good way, mabye the best way, to detemrine if cannabis will be good is to smell it. But this is difficult since in legal markets like California, pot is sold in preackaged containers. “Science Reverals the Cannabis industry’s greatest lie: you’re buyin g weed wrong (and so is everyone else), Chris Roberts, June 23, 2022). 

One cognitive test has shown that in fact there is no difference in cognition between the sober and stoned condition. (“Weed is tronger than ever, but it might not affect our brains that much: 6 surprising facts about cannabis” CC Documentaries.)

In another study, one group of participants were given flower buds with 16-24 % THC while extract users got oil with 70-90% THC levels. The reserchers closely monitored the participants’ blood, moods, and cognitive function. Surpirsingly, they found that potency did not track with intoxication levels. While there were striking differences in blood levels, they were similarly impaired. The results suggested that other components like cannabidiol or CBD, which is harvested from hemp or marijuana plants that contain less than 0.03% THC also is important. Tehre are also compounds called terpenes, which affect how cannabis influences one’s mind and body. All of these componets of the cannabis plant work together to give one a smooth and seamless high. Thus unlike gin or vodka, where alcohol levels define how drunk one gets, how potent your weed is can not be determiend by its THC content. 

Companies:  Texas Original   Good Blend

Cronos Group

Organizations: PBC Conference

Regulating Agencies: NY Office of Cannabis Management

See Also culturing techniques (outline)

Introduction/Definitions:

Cannabis is the genus of a variety of species—Cannabis sativa, Cannabis ruderalis, and Cannabis indica—which is often used as an umbrella term to refer to them all. This misclassification of the different species has made it difficult to properly distinguish between and understand the best ways to utilize the different varieties of these plants. (US PP34724 P2)

Cannabis has a lot of names and uses. The plant and its compoudns have fuelled markts from pre-rolled joints to gummies, soft drinks, vaporizers and oils. (Weed is tronger than ever, but it might not affect our brains that much: 6 surprising facts about cannabis” CC Documentaries. Cannbis was first domesticated in East Asia, which it grows naturaly. For about 4,000 years, human used its fibres to make hemp rope and clothing, and the plant’s oily seeds for food. Cannabis seeds have been found with Japanese pottery shards dated to about 10,000 years ago. It was traded along the Silk Road and spread across continents. (“Weed is tronger than ever, but it might not affect our brains that much: 6 surprising facts about cannabis” CC Documentaries.)

Cannabis, more commonly known as marijuana, is a genus of flowering plants that includes at least three species, Cannabis saliva, Cannabis indica, and Cannabis ruderalis as determined by plant phenotypes and secondary metabolite profiles. In practice however, cannabis nomenclature is often used incorrectly or interchangeable. Cannabis literature can be found referring to all cannabis varieties as “sativas” or all cannabinoid producing plants as “indicas”. Indeed the promiscuous crosses of indoor cannabis breeding programs have made it difficult to distinguish varieties, with most cannabis being sold in the United States having features of both sativa and indica species.

Cannabinoids: are compounds that can be found in the cannabis leaf, stem and bud. They have become widespread in products taht you can smoke, eat, vape, drink or rub on your skin, and are increasingly used to allevaite everything from pain to anxiety. Weed is tronger than ever, but it might not affect our brains that much: 6 surprising facts about cannabis” CC Documentaries.)

Cannabidiol (CBD): CBD which is also found in the cannabis plant has been shown to have medical benefits such as for the treatment of seizures.

THC in a cannabis bud can break down over time. When THC degrades, it converts to cannabinol.

Species of Cannabis:

Cannabis sativa L.: is a widespread species that is cultivated worldwise in wide-ranging habitates areas outside its natural range. It is considered one of the oldest domestic and cultivated plants in the history of mankind.

C. sativa L.. is a crop species that has multiple roles. It has been cultivated for industrial, nutritional, and medicinal purposes. For centries, ehmp stemps have been used for fibers (mats, shoes, cloth, and ropes) and its seeds have been sued for oil production. Moreover, hemp seeds are an excellent source of omega-3 and omega- fatty acids, as well as nother nutritious oil and proteins. Recently, the stem tissues have begun to be used in the mnaufacture of bioplastics and concrete-like material and for high-performance composition applications. (Mitsis, “An Alternative in vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) presenting efficient rooting, for commerical production” Plants, 2022, 11(10): 1333).

In recent decades, there has been a resurgence of interest in the use of bioactive compounds from natural sources, such as hemp, with constantly increasing demand. Hemp flowers primarily and its leaves incidentally produce about 545 bioactive secondary metabolites. The use of these substances, such as terpenoids, flavonoids, and phytosterols, and alkaloids and glycoproteins, as well as a special class of terpenophenolic compounds, the cannabinoids, is constantly increasing, adn the majroity of them have medicinal properties. Among the numberous cannabinoids existing in hemp flowers, the most studies phytocannabinoids in relation to their therapeutic uses are the intoxication delta-9-tetrahydrocannabinol (dalta9-THC), a rpomising medicinal compound for the treatment of various diseases with well-known medicinal effects; cannabidiol (CBD), which has several proven pharmacological propties and cannabigerol for its potential remedial effects. (Mitsis, “An Alternative in vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) presenting efficient rooting, for commerical production” Plants, 2022, 11(10): 1333).

Cannabinoid production through the exploitation of natural resources is constantly increasing, due to these plants’ apparent health, nutritional, and mostly medicinal properites. For instance, C. sativa L. sales in the UUS are expected to rise from 8 billion as recorded in 2018 to over 40 billion by 2025. This imposes a need to detect and preserve genetic resources of C. sativa L. varieties that are rich in bioactive secondary metabolites and to supply the international market with adequate quantities from reliable sources. (Mitsis, “An Alternative in vitro Propagation Protocol of Cannabis sativa L. (Cannabaceae) presenting efficient rooting, for commerical production” Plants, 2022, 11(10): 1333).

Genome of Cannabis:

Additional breeding methods have been known to one of ordinary skill in the art, e.g., methods discussed in Chahal and Gosal (Principles and procedures of plant breeding: biotechnological and conventional approaches, CRC Press, 2002, ISBN 084931321X, 9780849313219), Taji et al. (In vitro plant breeding, Routledge, 2002, ISBN 156022908X, 9781560229087), Richards (Plant breeding systems, Taylor & Francis US, 1997, ISBN 0412574500, 9780412574504), Hayes (Methods of Plant Breeding, Publisher: READ BOOKS, 2007, ISBN1406737062, 9781406737066), each of which is incorporated by reference in its entirety for all purposes. Cannabis genome has been sequenced (Bakel et al., The draft genome and transcriptome of Cannabis sativa, Genome Biology, 12 (10):R102, 2011). Molecular markers for Cannabis plants are described in Datwyler et al. (Genetic variation in hemp and marijuana ( Cannabis sativa L.) according to amplified fragment length polymorphisms, J Forensic Sci. 2006 March; 51 (2):371-5), Pinarkara et al., (RAPD analysis of seized marijuana ( Cannabis sativa L.) in Turkey, Electronic Journal of Biotechnology, 12 (1), 2009), Hakki et al., (Inter simple sequence repeats separate efficiently hemp from marijuana ( Cannabis sativa L.), Electronic Journal of Biotechnology, 10 (4), 2007), Datwyler et al., (Genetic Variation in Hemp and Marijuana ( Cannabis sativa L.) According to Amplified Fragment Length Polymorphisms, J Forensic Sci, March 2006, 51 (2):371-375), Gilmore et al. (Isolation of microsatellite markers in Cannabis sativa L. (marijuana), Molecular Ecology Notes, 3 (1):105-107, March 2003), Pacifico et al., (Genetics and marker-assisted selection of chemotype in Cannabis sativa L.), Molecular Breeding (2006) 17:257-268), and Mendoza et al., (Genetic individualization of Cannabis sativa by a short tandem repeat multiplex system, Anal Bioanal Chem (2009) 393:719-726), each of which is herein incorporated by reference in its entirety for all purposes.

In 2011 , researchers at the Unviersity of Saskatchewan sequenced the genome of Cannabis sativa. They discovered that over thousands of years, human selectively bred the plant into two strains: one for fiberes and seeds, and one for medicine. The anlysis showed that the synthase gene of THC’s preducrsor, THCA, an essential enzme in THCA production, is turned on in marijuana, but switched off in hemp. Hemp has been widely used for millennia. Besides rope and clothing, its been sued for everything from art canvases to sails to paper. More recently, hempcrete, a durale, lightweight composit reinforced with hemp, has been used in construction nd can replace materials like drywal, insulation and siding. (“Weed is tronger than ever, but it might not affect our brains that much: 6 surprising facts about cannabis” CC Documentaries.)

Legalization of Cannabis:

See also T cell activaiton in immune cells

Antigen recognition by B cells is mediated by the B cell receptor (BCR), a surface-bound immunoglobulin in complex with signaling componets CD79alpha and CD79beta. Crosslinking of BCR upon engagement of antigen results in phosphorylation of immunoreceptor tyrosin-based activation motifs (ITAMs) within CD79 a and CD79b, initiating a cascade of intracellular signaling events that recruits downstream molecules to the membrane and stimulate calcium mobilization. This leads to the induction of diverse B cell responses (e.g., cell suvival, proliferation, antibody production, antigen presentation, differentiation, etc) which lead to a humoral iimmune response. Other components of the BCR coreceptor complex ehnace (e.g., CD19, CD21, and CD81) or suppress (e.g., CD22 and CD72) BCR activation signals. (Chu, US 2012/0321620).

Negative feedback of BCR

Antibody negative feedback

When antibodies are produed to an antigen, the circulating level of immune complexes (e.g., antigen bound to antibody) increases. These imune complexes downregulate antigen-induced B cell activation. It is believed that these immune complexes downregulate antigen-induced B cell activaiton by coengaging cognate BCR with the low-affinity inhibitory receptor Fc.gamma.RIIb, the only IgG receptor on the B cells. It is also beleived that this negative feedback of antibody production requires interaction of the antibody Fc domain with FcgammaRIIb since immune complexes containing (F(ab’).sub.2 antibody fragmetns are not inhibitory. The intracellular immunoreceptor tyrosine-based inhibitory. motif (ITIM) of Fc.gamma.RIIb is necessary to inhibit BCR induced intracellular signals. This inhibitory effet occurs through phosphorylation of teh Fc.gamma.RIIb ITIM, which recurits SH2-containing inositol polyphosphate 5-phosphatase (SHIP) to neutralize ITAM-induced intracellular calcium mobilization. In addition, Fc.gamma.RIIb-mediated SHIP phosphorylation inhibits the downstream Ras-MAP proliferation patway.. (Chu, US 2012/0321620). 

Chu, US 2012/0321620) disclosees immunoglobulins that inhibit cells that ewxpress Fc.gamma.RIIb. The antibodies binds Fc.gamma.RIIb and coengages a target antigen on the cell’s surface and coengages a target antigen on the cell’s surface and an Fc.gamma.RIIb on the cell’s surface. 

See also MAP Kinases

Classes and Structure:

The PI3Ks are an ancient family of intracellular kinases that evolved to mediate nutrient sensing and metabolic control. In mammals, there are 8 different PI3K catlytic subunits, dviided into three classes. Class I PI3Ks phosphorylate phosphatidylinositol (,5)P2 (PIP2) to geenrate phosphatidylinositol (3,4,5)P3 (PIP3) which acts as pivotal second messenger signaling moleule. In B cells, both AKt and Btk can bind to PIP3 via their PH domains. PIP3 is essential for teh activaiton of Akt and contributes to the activation of Btk. Less is known about the role of classes II and III PI3Ks in B cells. (Okkenhaug, “PI3k signaling in normal B cells and chronic lymphocytic leukemia”, Current Topics in Microbiology and Immunology )2106) 393: 123-142). 

Class I phosphoinositide 3-kinases (PI 3-kinases; PI3Ks):

Class I PI3Ks are of particualr interest and are futher divided into class IA and class IB kinases based on sequence homology and substrate specificity. Class IA PI3Ks contain a p85 regulatory subunit that therodimerizes with a p100, alpha, beta or delta catalytic subunit. These kinases are commonly known as PI3Kalpha, beta and delta respectiviely and are activated by receptor tyrosinekinases. The class IB PI3k contains a p110gamma ctalyti subunit and is commonly known as PI3kgamma which is activated by heterotrimeric G proteins. (Qiao, US 2012/0258967). 

Mammals, have 4 differnt class I PI3Ks. Heterodimers of a regultory subunit (p85alpha, p55alpha, p50alpha, p85beta or p55gamme, collectively referred to as p85) and a catlytic subunit (p110alpha, p110beta or p110epsilon) form PI3kalpha, PI3kbeta or PI3kbeta wehreas PI3kgamma is a hterodimer of p101 or p84 with p110gamma. (Okkenhaug, “PI3k signaling in normal B cells and chronic lymphocytic leukemia”, Current Topics in Microbiology and Immunology )2106) 393: 123-142).

Structurally, class I PI3Ks exists as heterodimeric complexes, consisting of a p100 catalytic subunit and a p55, p85, or p101 regulatory subunit. There are four p110 catalytic subunits. Class I PI3Ks can be further divided into two subclasses (Ia and Ib) based on ther mechanism of action. 

PI3 kinase is classified into three classes (iel, classes I-III) in terms of structure. Class I is further divided into classes IA adn 1B and of these, class IA is divided into three isotypes, p110alpha, p110beta adn p110epsilon  (Niikura, US Patent Application No: 16/484,061, published as US 2019/0359691). 

Functions:

PI3 Kinase (phosphoinositide 3-kinase: PI3K) phosphorylates the position 3 of the inositol ring of phosphatidylinositol, and plays an important role in various cellular functions such as cell survival, cell growth, cell motility, and the transport of intracellular organelle.  (Niikura, US Patent Application No: 16/484,061, published as US 2019/0359691). 

Antibody diversity: Niikura, *US Patent Application No: 16/484,061, published as US 2019/0359691) discloses a process for promoting diversificaiton of amino acid sequences of variable regions of an antibody generated by an avian B cell poluation that includes suppressing PI3Kalpha activity of an avian b cell population. In one embodiment, the PI3kalpha-specific inhibitor is PI3Kalpha inhibitor 2 or A66. 

Antibody Class Switching: PI3 Kinase (phosphoinositide 3-kinase, PI3k) has been known as a factor functioning upstream of AID. As a result of studies using a PI3K inhibitor in mouse B cells, it has been demonstrated that when p110episolon signaling is suppressed, the expression of AID is increased and class switch recombination is promoted. (Niikura, US Patent Application No: 16/484,061, published as US 2019/0359691). 

Role in Leukocyte chemotaxis: PI3Ks are known to play a pivotal role in the ability of leukocytes to undergo chemotaxis as the lipid products they generate, including but not limited to phosphatidylinositol (3,4,5)-trisphosphate (PI3), are ciritical for promoting asymmetric F-actin synthesis, and thus luekocyte cell polization. The function of class I PI3Ks, however, is not limited to directed migration, in that they are also required for phagocytosis and generation of oxygen radicals in response to chemoattractants. The ability of class I PI3Ks to regulated these processes in leukocytes relies on PIP3 mediated recruitment of two lipd-binding protein inases, phosphatidylinositol-dependent kinase 1 (PDK1) and protein kinase B/Akt, both of which can interact with this PI-derivative via their pleckstrin homology domains. Association of these kinases with PIP3 at the plasma membrane brings them into close proximity, facilitating the phosphorylation and activaiton of Akt by PDK1. These proteins are, in turn, responsible for many of the donstream signaling events associated with PI3K activity.

Role in neutrophil cell migration: Evidence supporting the class I PI3Ks involvement in neutrophil cell migration is found in the ability of non-selective class I PI3k inhibitors to mitigate neutrophil chemotaxis.

Effect on TLR3 Signalling: It is known that PI3 kinase activation is required for TLR3-induced IRF3 activation and subsequent IFN-beta gene induction.

Inhibitors of PI3 Kinase: 

LY294002 is a pharmacological inibitor of PI#k.

PKC activity has been coupled with the preservation of cell survival. The catechin EGCG has been reported to protect neuronal cell lines against cell death induced by 24 h exposure to 6-OHDA.

Inhibitors of PKC:

GF109203X is a general PKC inhibitor. Prior exposure of nueronal cells to this inhibitor completely abolished the protective effect of EGCG against cell death.

Given the important role that NF-kB plays in the regulation of a large number of proinflammatory genes, there is a growing interest in targeting it in order to affect the inherent redundancy of the inflammatory cascade. A large body of evidence links the NF-kB pathway to the dysregulated inflammation that is characteristic of diseases such as sepsis and acute respiratory distress syndrome. Several of the genes that comprise the complex network contributing to this dysregulated inflammation and are regulated at the transcriptional level by NF-kB, including the cytokines IL-1? and TNF-alpha, chemokines such as IL-6, IL-8 and macrophage chemotactic protein-1; cell adhesion molecules such as vascular cell adhesion molecule 1 and intercellular adhesion molecule 1; growth factors such as granulocyte macrophage colony-stimulating factor and granulocyte colony stimulating factor; as well as additional proinflammatory genes such as inducible nitric oxide synthase. 

Agents Known to Inhibit NF-kB: Pharmacoglogical agents inhibit NF-kB at one of its activation steps, and several classes of drugs are well-known NF-kB inhibitors. such agents include the following: See also 

glucocorticoids such as dexamethasone and prednisone

immunosuppressants such as cyclosporine, tacrolimus, and deoxyspergualin

nonsteroidal anti-inflammatory drugs such as aspirin, sodium slicylate and tepoxalin.

capsaicin

Helenalin: is an inhibitor of NF-kB

hypericin

epigallocatechin-e-gallate (EGCG) which is the major polyphenol present in green tea is a potential novel, safe, and nontoxic strategy for inhibiting NF-kB. 

SN50

Bacterial toxins or virulence factors: 

Microbes interfere with the activation of NF-kB at various parts of the signaling pathway. First part these pathways can inhibit is the pathway proximal to the phosphorylation and degradation of IkB. An example of this is provided by vaccinia virus, which produces a viral homologue of MyK88 which antagonizes MyD88 dependent activaiton of NF-kB. 

The ability to intefere with the degradation of IkB is another strategy employed by some pathogens. For example, the measles virus is able to prevent phosphorylation of IkBalpha in nueronal cells. 

Some pathogens are able to interfere with NFkB activation downstream from the degradation of IkB. For example, the African swin fever virus makes a viral protein which is a homologue of IkB which can bind to NFkB following degradation of host IkB and so inhibits nuclear localization of dimers. 

Some pathgoenic bacteria also interfere with the ability of NFkB dimers to translocate into the nucleus as by perhaps by directly interfering with the ability of NF-kB to bind DNA. 

Yersinia and Salmonella inhibit NF-kB activation.

antrax bacterium induces apoptosis of TLR4 activated macrophages through its lethal toxin, whose lethal factor catalytic subunit prevetns p38 activaiton by cleavage of the upstream kinase MKK6 (Park et al., 2002) 

Negative Regulation of NF-kB: The IkB-alpha gene (a subtype of the class of IkB inhibitors and clearly taregeted in the case of inflammatory stimulation) also contains an NF-kB-responsive element in its promoter which leads to upregulation by NF-kB and to resynthesis of the inhibitor molecule which can now catch and binding newly synthesised as well as previously released NF-kB. This acts as an autoregulatory loop (negative feedback) mechanism, keeping the NF-kB levels in balance and gene transcription under tight control.

All known  have been shown to signal through Toll/IL-1R homology (TIR) domain-containing adaptor molecules, MyD88, and/or TIR domain-containing adaptor-inducing IFN-? (TRIF). Mice deficient in both MyD88 and TRIF are unresponsive to stimulation with all known TLR ligands. 

TNF?:

TNF blockagde is very effective in the treatment of rheumatoid arthritis. The reason that TNF is so imiportant is not entirely clear. However, TNF signals through an evolutionarily conserved pathway and is a point of connection between two arms of the ammalian inante immune response. It is a second wave activator of NF-kB, capable of spreading the alarm sounded by TLRs to virtually all cell types in teh body.

Upon TNF? binding to the TNF receptor, the receptors aggregate and bind adaptor proteins, leading to activation of the IkB kinase (IKK) complex. Phosphorylation of IkB by IKK leads to ubiquitination and degradation of IkB and allows free NF-kB to bind target genes. one such target is IkB?, and its production results in a negative feedback loop.

LPS:

LPS signals through TLR4. TLR4 activates two downstream pathways. a MyD88-dependent pathway that leads to the production of proinflammatory cytokines such as IL-6, TNF and IL-12 with quick activation of NF-kB and MAPK and a MyD88-independent pathway associated with activation of IRF-3, subsequent induction of IFN-beta, and maturation of DCs, with delayed activation of NF-kB and MAPK. Thus each pathway is thought to directly activate NF-kB. The MyD88 dependent pathways involves the early phase of NFkB activation which leads to the production of inflammatory cytokines. The MyD88 independent pathway activates interferon (IFN) regulatory factor (IRF3) and involves the late phase of NFkB activation, both of which lead to the produciton of IFNb and the expression of IFN inducible genes.

1. The MyD88-dependent pathway recruits the kinases interleukin-1 receptor-associated kinase 1 (IRAK1) and IRAK4, which phosphorylate TNF receptor associated factor 6 (TRAF6), leading to the activation of the IKK complex. NF-kB activation through the MyD88 dependent pathway occurs earlier than through the MyD88 independent pathway. MyD88 intereacts with TLR4 thourgh a TIR domain and recruits IRAK to the receptor cojmplex through a death domain, and then IRAK is sequentially activated. Activated IRAK subsequently activates NFkB via TNFR-associated factor (TRAF).

2. MyD88-independent pathway: is not completely understood. The pathway is dependent on the TIR domain-containing adaptor inducing interferon-?(Trif) adaptor molecule, and Trif-related adaptor molecule (Tram), receptor-interactor protein 1 (RIP1), and RIP3 have been identified as important factors in the pathway. In addition to inducing expression of IFN-inducible genes, the MyD88 independent pathway leads to the LPS mediated maturation of DCs. When cultured with LPS, MyD88 deficient BMDCs upregulate the cell surface expression of co-stimulatory molecules, such as CD40, CD80 and CD86 and induce the proliferation of T cells. By contrast, TLR4 defiicent DCs fail to mature in response to LPS, indicating that DC maturation proceeds in a MyD88 independent manner. 

Interferon-regulatory factor 3 (IRF3) is a MyD88 independent pathway specific factor that directly regulates early response genes (for example, those encoding interferon-? and IP-10) and is active within 30 min of LPS stimulation. 

The end result of these two pathways is the same as with the TNF? activated pathway: degradation of IkB, which is followed by activation of IkB?gene transcription.

 From Nature, 430 (2004)

From: Science, 26 September 2005, vol 309