The short version of Freeze-thaw fits in a sentence. The long version — which is the one that helps — is below.
This page was last updated on 2026-05-21 and is reviewed periodically as new material appears.
NAD+ is commonly measured by high-performance liquid chromatography with ultraviolet detection, often at 254 or 260 nm. Enzymatic cycling assays provide higher sensitivity by coupling NAD+ to a reporter reaction. Mass spectrometry can distinguish NAD+ from close analogues and confirm isotope labeling. Sample preparation usually involves rapid quenching of metabolism to prevent interconversion with NADH. Because NAD+ and NADH differ by one hydride, extraction conditions strongly affect the measured ratio.
In aqueous solution, NAD+ is most stable under mildly acidic to neutral conditions and degrades faster at high pH or elevated temperature. The molecule can hydrolyze at the pyrophosphate bond or undergo nonenzymatic cyclization. Buffers, chelating agents, and cold temperatures slow these losses during analysis. Repeated freeze-thaw cycles are generally avoided because they can promote degradation and concentration changes. Light exposure is also controlled, though NAD+ is less photolabile than some related nucleotides.
Commercial NAD+ is supplied as a solid, often as the free acid or a salt, and purity is verified by chromatographic methods. Laboratories typically store it desiccated at minus 20 degrees Celsius or below. Working solutions are prepared fresh because even sterile aqueous solutions can lose activity over hours to days depending on pH and temperature. Documentation may include a certificate of analysis, an assay value, and a recommended retest date. Researchers should verify identity and purity when results depend on precise cofactor concentrations.
The molecule was first described in the early twentieth century as a factor that promoted fermentation in yeast extracts. Later work linked it to hydrogen transfer and to the oxidation of nutrients in living tissues. Its structure was resolved as a dinucleotide, which explained why it could accept and donate electrons at specific enzyme sites. Today, NAD+ is recognized as a central substrate and signaling precursor, not merely a metabolic cofactor. Whether all observed NAD+ changes reflect causal signaling remains an open question.
Related compounds include NADH, the reduced form, and NADP+, which carries an additional phosphate group. NADP+ and NADPH often serve in biosynthetic and antioxidant reactions, while NAD+ and NADH are more associated with energy-yielding catabolism. Nicotinamide, nicotinic acid, and nicotinamide riboside are precursors that can enter salvage pathways. The exact contribution of dietary precursors to tissue NAD+ pools is an area of active investigation. Some studies measure labeled precursors to trace those routes.
NAD+ is the oxidized form of nicotinamide adenine dinucleotide, a coenzyme built from two nucleotides joined by a phosphate linkage. One nucleotide carries adenine, and the other carries nicotinamide; the plus sign denotes a formal positive charge on the nicotinamide ring, not a free proton. In cells, NAD+ and its reduced partner NADH form a reversible redox pair. That pair participates in electron transfer reactions throughout metabolism. The abbreviation NAD+ is common in biochemistry, while NAD(H) sometimes denotes the combined pool.
| Property | Value | Notes |
|---|---|---|
| UV absorption maximum | 259–260 nm | Aqueous solution; pH-dependent |
| Common salt form | Disodium salt | Improves aqueous solubility |
| Typical storage temperature | -20 °C or lower | Desiccated and protected from light |
| Common analytical method | HPLC with UV detection | Often paired with mass spectrometry |
| Aqueous stability | pH and temperature dependent | Degrades faster at alkaline pH and high heat |
Nicotinamide adenine dinucleotide, commonly abbreviated NAD+, is a coenzyme present in all living cells. The molecule consists of two nucleotides linked by phosphate groups, with adenine and a nicotinamide ring as its principal features. In its oxidized form, the nicotinamide ring can accept a hydride ion, becoming NADH. This reversible conversion places NAD+ at the center of many electron-transfer reactions. Its role as a redox carrier is well established across bacteria, plants, fungi, and animals.
Beyond redox chemistry, NAD+ acts as a substrate for several enzyme families. ADP-ribosyltransferases, sirtuins, and CD38 ectoenzymes cleave the molecule into nicotinamide and ADP-ribose or related products. These reactions connect NAD+ availability to processes such as DNA repair, chromatin modification, and calcium signaling. Because the coenzyme is used in both electron transfer and signaling, cells maintain separate pools in compartments including the cytosol, mitochondria, and nucleus. The relative sizes and regulation of those pools remain active areas of study.
Cells produce NAD+ through several biosynthetic routes. The salvage pathway recycles nicotinamide, while the Preiss-Handler pathway uses nicotinic acid, and a de novo route can start from tryptophan in some organisms. In mammals, the salvage pathway is generally considered the main source under ordinary conditions. Tissue concentrations vary widely by cell type and compartment, and measured declines with age have been reported in some studies. Whether such changes drive aging or mainly accompany it remains an open question.
In redox reactions, NAD+ accepts a hydride ion, which consists of two electrons and one proton. The hydride adds to the nicotinamide ring at a specific carbon, converting NAD+ into NADH. Dehydrogenase enzymes use this step in glycolysis, the citric acid cycle, and fatty acid oxidation. NADH later donates electrons to the mitochondrial electron transport chain, helping to drive ATP synthesis. The balance between NAD+ and NADH reflects the metabolic state of a cell, and shifts in that balance can alter how pathways operate.
Beyond electron transfer, NAD+ serves as a substrate for enzymes that cleave it and attach its ADP-ribose portion to other molecules. This group includes poly(ADP-ribose) polymerases, CD38, and sirtuins. Such reactions consume NAD+ and can influence its availability for metabolism. Cells replenish NAD+ through a salvage pathway that recycles nicotinamide and through routes starting from tryptophan or vitamin B3 forms. How these synthesis and consumption routes are coordinated across tissues remains an active area of study, and compartment-specific concentrations are difficult to measure directly.
Nicotinamide adenine dinucleotide, abbreviated NAD+, is a dinucleotide built from adenine, nicotinamide, two ribose sugars, and two phosphate groups. The oxidized form carries a positive charge on the nicotinamide ring, while the reduced form, NADH, is neutral. This pair acts as a reversible electron carrier in cells. NAD+ is present in bacteria, plants, animals, and fungi. Its structure allows it to accept and donate electrons without being consumed in the reactions it supports.
The same procedure was followed as in Annis, but the sample of galaxies used is 80 times larger than that used in the Annis study. Some sources were classified as "lopsided": they appear asymmetric in shape, meaning that one side of the galactic disc is more massive and less luminous than the other. This characteristic, according to Calissendorff, could be an indication that the galaxy is home to a civilization that has placed Dyson spheres in its main part. This can be explained by the fact that the colonization starts from one side of the galactic disk, making it appear darker and leading a distant observer to believe that the core has moved to that same side. On the other hand, a galaxy hosting Dyson spheres should be characterized by a significant source of far-infrared radiation. The fact remains that a Type III civilization can consume energy through a Dyson sphere without surrounding a star. Indeed, such megastructures could also extract energy from a black hole, according to the study by Inoue and Yokoo (2011). However, such a structure would not reduce the luminosity of an observed galaxy. Calissendorff's study concludes that 11 of the sources analyzed (out of a catalog of 2,411 galaxies, or 0.46%) show possible evidence of a Type III civilization. Searching for objects that obscure 90% of the light leaves only one source remains that meets the criteria. These positive sources show a low redshift (so they are old, about 100 million years), which is consistent with possible Type III civilizations, that could have flourished only in the early past.
=== COVID-19 misinformation === During the COVID-19 pandemic, Ayyadurai used social media to spread various conspiracy theories and misinformation about the pandemic. In January 2020, he claimed that COVID-19 was patented by the Pirbright Institute, but the patent he referenced relates to avian coronavirus, which infects birds, not SARS-CoV-2, the virus responsible for the pandemic. Ayyadurai defined COVID-19 as "an overactive dysfunctional immune system that overreacts and that's what causes damage to the body", and claimed that vitamin C could be used to treat it. He alleged that COVID-19 was spread by the "deep state" and accused Anthony Fauci, director of the National Institute of Allergy and Infectious Diseases, of being a "Deep State Plant". Ayyadurai called for Fauci to be fired and his supporters lobbied for Fauci to be replaced by Ayyadurai. In March 2020, Ayyadurai published an open letter to then-U.S. President Donald Trump, writing that a national lockdown was unnecessary and advocated that large doses of vitamins could prevent and cure COVID-19. In April 2020, Politico and Vanity Fair reported that QAnon supporter DeAnna Lorraine recommended that Ayyadurai be included in COVID-19 discussions at Donald Trump's White House.
=== Chemicals === E-cigarette components include a mouthpiece, a cartridge (liquid storage area), a heating element or atomizer, a microprocessor, a battery, and some of them have a LED light at the tip. They are disposable or reusable devices. Disposable ones are not rechargeable and typically cannot be refilled with a liquid. There is a diverse range of disposable and reusable devices, resulting in broad variations in their structure and their performance. Since many devices include interchangeable components, users have the ability to alter the nature of the inhaled vapor. For the majority of e-cigarettes, many aspects are similar to their traditional counterparts, such as giving nicotine to the user. The use of an e-cigarette simulates the action of smoking, with a vapor that somewhat resembles cigarette smoke. E-cigarettes do not involve tobacco combustion, and they do not produce vapor between puffs. They do not produce sidestream smoke or sidestream vapor. Vapor production basically entails preprocessing, vapor generation, and postprocessing. First, the e-cigarette is activated by pressing a button, or other devices switch on by an airflow sensor or other type of trigger sensor. Then, power is released to an LED, other sensors, and other parts of the device, and to a heating element or other kind of vapor generator. Subsequently, the e-liquid flows by capillary action to the heating element or other devices to the e-cigarette vapor generator. Second, the e-cigarette vapor processing entails vapor generation.
deoxyribose Also 2-deoxyribose. A monosaccharide pentose sugar derived from ribose by the replacement of the hydroxyl group attached to the C2 carbon with a single hydrogen atom. D-deoxyribose, in its cyclic ring form, is one of three main functional groups of deoxyribonucleotides and hence of deoxyribonucleic acid (DNA) molecules.
Sources: en.wikipedia.org
Surgery is an ancient medical specialty that uses operative manual and instrumental techniques on a patient to investigate or treat a pathological condition such as disease or injury, to help improve bodily function or appearance or to repair unwanted ruptured areas (for example, a perforated ear drum). Surgeons must also manage pre-operative, post-operative, and potential surgical candidates on the hospital wards. In some centers, anesthesiology is part of the division of surgery (for historical and logistical reasons), although it is not a surgical discipline. Other medical specialties may employ surgical procedures, such as ophthalmology and dermatology, but are not considered surgical sub-specialties per se. Surgical training in the U.S. requires a minimum of five years of residency after medical school. Sub-specialties of surgery often require seven or more years. In addition, fellowships can last an additional one to three years. Because post-residency fellowships can be competitive, many trainees devote two additional years to research. Thus in some cases surgical training will not finish until more than a decade after medical school. Furthermore, surgical training can be very difficult and time-consuming. Surgical subspecialties include those a physician may specialize in after undergoing general surgery residency training as well as several surgical fields with separate residency training. Surgical subspecialties that one may pursue following general surgery residency training:
Various strategies have been employed for constraining α-helices, including the non-covalent and covalent stabilization techniques; however, the all-hydrocarbon covalent link, termed a peptide staple, has been shown to have improved stability and cell penetrability, making this stabilization strategy particularly relevant for clinical applications.
Diazepam is a medication with a high risk of misuse and can cause drug dependence. Some pharmacologists recommend urgent action by national governments to improve prescribing patterns of benzodiazepines such as diazepam. A single dose of diazepam modulates the dopamine system in similar ways to how morphine and alcohol modulate the dopaminergic pathways. Between 50 and 64% of rats will self-administer diazepam. Diazepam can substitute for the behavioral effects of barbiturates in a primate study. Diazepam has been found as an adulterant in heroin. Diazepam drug misuse can occur either through recreational misuse, where the drug is taken to achieve a high, or when the drug is continued long term against medical advice. Sometimes, it is used by stimulant users to "come down" and sleep and to help control the urge to binge. These users often escalate dosage from 2 to 25 times the therapeutic dose of 5 mg to 10 mg. A large-scale study in the US, conducted by SAMHSA, using data from 2011, determined that benzodiazepines were present in 28.7% of emergency department visits involving nonmedical use of pharmaceuticals. In this regard, benzodiazepines are second only to opiates, the study found in 39.2% of visits. About 29.3% of drug-related suicide attempts involve benzodiazepines, making them the most frequently represented class in drug-related suicide attempts. Males misuse benzodiazepines as commonly as females.
Sources: en.wikipedia.org
=== Treatment === Gastrointestinal surgery can often be performed in the outpatient setting. In the United States in 2012, operations on the digestive system accounted for 3 of the 25 most common ambulatory surgery procedures and constituted 9.1 percent of all outpatient ambulatory surgeries.
Furthermore, the marked brocho-constriction produced by injections of histamine was not reversed by either l-synephrine or d,l-synephrine. In experiments with isolated sheep carotid artery, d-, l- and d,l-synephrine all showed some vasoconstrictor activity: l-synephrine was the most potent, producing strong contractions at a concentration of 1:10000. d-Synephrine was about 1/2 as potent as the l-isomer, but d,l-synephrine (which would have been expected to have a potency of 1/2 that of l-synephrine even if the d-isomer were completely inactive) did not produce significant and irregular contractions until a concentration of 1:2500had been reached, implying an inhibitory interaction between the two enantiomers. Qualitatively similar results were obtained in a rabbit ear preparation: 25 mg l-synephrine produced significant (50%) vasoconstriction, while the same concentration of d-synephrine elicited essentially no response. In contrast, d,l-synephrine did not produce any constriction up to 25 mg, but 25 – 50 mg caused a relaxation of the blood vessels, which again suggested that the d-isomer might be inhibiting the action of the l-isomer. Experiments on strips of rabbit duodenum showed that l-synephrine caused a modest reduction in contractions at a concentration of 1:17000, but that the effects of the d- and d,l- forms were much weaker. Racemic synephrine, given intramuscularly, or by instillation, was found to significantly reduce the inflammation caused by instillation of mustard oil into the eyes of rabbits.
==== Memorial ceremony for the Martyrs of San Juan de Wilson ==== At 6:30pm on March 5, 1870, San Andrés Lodge No. 3, in Havana, held a memorial ceremony to observe Masonic funeral rites for the deceased Grand Master Andrés Puente. Despite the fact that certain members of the Lodge were afraid that a memorial gathering would put their lives in danger, nevertheless more than 100 people made a fully-packed temple that night, with all Lodge members present. The local police, however, had also known that there would be a memorial that night. When the temple was full, the armed and uniformed police mustered in the street out front of the Lodge. As the police were getting into formation, local citizens gathered behind them and grew into a large mob, chanting and crying out: "Kill the conspirators, death to the Cuban Council!" When the police breached the building, the Outside Guard informed Worshipful Master Nicolas Dominguez Cowan that the police were on the stairs. Cowan addressed the Freemasons: "Do not be alarmed, my dear brethren, our fate, whatever it may be, we must face with courage; we must save our charter above everything else." Edward Godwin, a government employee at the local customs house, escaped the Lodge with the charter and brought it to the safety at a house belonging to another Freemason, where he stashed it. Of the over 100 people in the building that night, over 40 of them were arrested and taken to prison. Some of the members were able to evade capture. Those forty or more Freemasons who were arrested that night were imprisoned for one hundred days.
Injury in animals is sometimes defined as mechanical damage to an anatomical structure, but it has a wider connotation of physical damage with any cause, including drowning, burns, and poisoning. Such damage may result from attempted predation, territorial fights, falls, and abiotic factors. Injury triggers an inflammatory response in animals of many different phyla. This prompts coagulation of the blood or body fluid, followed by wound healing, which may be rapid, as in cnidarians. Arthropods are able to repair injuries to the cuticle that forms their exoskeleton to some extent. Animals in several phyla, including annelids, arthropods, cnidarians, molluscs, nematodes, and vertebrates, are able to produce antimicrobial peptides to fight off infection following an injury.
Sources: en.wikipedia.org
NAD+ and NADH can interconvert quickly after a sample is collected, which can alter the measured ratio. Rapid quenching and cold handling limit enzymatic and chemical changes.
Purity is often checked by HPLC with UV detection, sometimes paired with mass spectrometry for identity. An assay against a standard can quantify the active cofactor content.
Solid NAD+ is usually kept dry, cold, and protected from light. Aqueous working solutions are best prepared fresh because degradation depends on pH, temperature, and time.
It indicates a formal positive charge on the nicotinamide ring. The molecule is not simply a protonated acid, and the charge is part of its redox chemistry.